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ImmunoTools cd4-fitc-(immunotools) antibody
Cd4 Fitc (Immunotools) Antibody, supplied by ImmunoTools, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cd4-fitc-(immunotools)+antibody/cd4+fitc++immunotools++antibody/us07807642-762-14-11
Average 90 stars, based on 1 article reviews
cd4-fitc-(immunotools) antibody - by Bioz Stars, 2026-11
90/100 stars

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Centrifugation:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Staining:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Flow Cytometry:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Cell Culture:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Isolation:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Inhibition:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Purification:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Dilution Assay:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Incubation:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Expressing:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Co-Culture Assay:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Variant Assay:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Microscopy:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

Enzyme-linked Immunosorbent Assay:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).

MANN-WHITNEY:

Article Title: Treg cell epitopes from α -tubulin: discovery and immunomodulatory features
Article Snippet: Finally, cells were washed with PBS by centrifugation at 300 g for 5 minutes and surface stained with antibodies anti-CD3 (HIT3a, PE/Cyanine5, Biolegend), anti-CD4 (MEM-241, FITC, Immunotools) and anti-CD25 (MEM-181, APC, Immunotools).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse anti-human CD28 (CD28.2, #555726, 2 μg mL −1 ), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL −1 ), goat anti-mouse (GAM, # 553998, 2 μg mL −1 ), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: Binding of Staphylococcal Enterotoxin B (SEB) to B7 Receptors Triggers TCR- and CD28-Mediated Inflammatory Signals in the Absence of MHC Class II Molecules
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803), anti-human CD86-FITC (#21480863), anti-human CD4-PE (#21278044), anti-human CD4-FITC (#21850043), anti-human CD8-APC (#21620086) (Immunotools, Germany); anti-human CD8-FITC (#555634), anti-human CD28-FITC (#556621), anti-human CD86-PE (#555658), anti-human CD3-PE (#555333) (BD Biosciences, Italy); anti-human HLA-DR-APC (#130-095-297), anti-human CD28-PE (#130-109-441) (Miltenyi Biotec, Italy); superagonistic anti-human CD28 (ANC28.1, #177820) (Ancell, USA); rabbit anti-GAPDH (#sc-25778), rabbit anti-HA (Y11, #sc-805) (Santa Cruz Biotechnology, USA); sheep anti-SLP76 (#06-548), mouse anti-pTyr (#05-321, Merck-Millipore, Italy); mouse anti-human CD3 (UCHT1, #555330), goat anti-mouse Alexa-flour 594 (#A11020), phalloidin-633 (#A22284) (ThermoFisher Scientific, Italy).

Article Title: Analysis of Surface Levels of IL-1 Receptors and Macrophage Scavenger Receptor I in Peripheral Immune Cells of Patients With Alzheimer Disease
Article Snippet: Staining was performed for 30 minutes at 4°C with fluorochrome-conjugated antibodies CD4-FITC (clone EDU-2), CD8-PE (clone MEM-31), CD14-PECy5 (clone 18D11), CD19-APC (clone LT-19; all Immunotools, Friesoythe, Germany), MSRI-PE (clone 351615), IL-1RII-FITC (clone 34141), and IL-1RI-PE (FAB269; all R&D Systems, Minneapolis).

Article Title: Peripheral differentiation patterns of human T cells
Article Snippet: For staining of surface antigens, cells were incubated 30 min in +4°C with anti‐human fluorescent‐labeled antibodies (CD8‐PE; clone MEM‐31, CD14‐FITC; clone 18D11, CD19‐FITC; clone Hi19a, and CD27‐APC; clone LT27 from Immunotools, Germany, and CD3‐APC‐H7; clone SK7, CD4‐BV510; clone SK3, CD21‐BV650; clone B‐ly4, CD28‐AlexaFluor700; clone CD28.2, CD31‐PeCy7; clone WM59, CD45RA‐BV711; clone HI100, CD95(FAS)‐BV421; clone DX2, CCR7‐PeCF594; clone 150503 from BD Biosciences, USA) and with Live/dead Fixable Green Dead Cell Stain (ThermoFisher Scientific, USA) that were diluted in Brilliant Stain Buffer (BD Biosciences).

Article Title: Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR
Article Snippet: Both the fresh CB and thawed CB segments were prepared by first lysing the red blood cells (RBC) with BD Pharm Lyse (BD Pharmingen, Berkshire, UK) before surface antibody staining at 4°C with CD4-FITC (ImmunoTools, Friesoythe; Germany; MEM-241), CD127-PE (BD Pharmingen; HIL-7R-M21), CD25-APC (BD Biosciences; Berkshire, UK; 2A3), CD3-PE-Cy7 (BD Pharmingen; SK7), and CD45-Alexa Fluor 700 (BD Pharmingen; HI30).

Article Title: SARS-CoV-2 Spike Does Not Possess Intrinsic Superantigen-like Inflammatory Activity.
Article Snippet: The following antibodies were used: anti-human CD80-FITC (#21270803, 1:10 dilution), anti-human CD4-FITC (#21278043, 1:10 dilution), anti-human CD86-PE (#555658, 1:10 dilution) (ImmunoTools, Germany); anti-human CD28-PE (# 130-109-441, 1:10 dilution), anti-human CD4-PE (#130-091-231, 1:10 dilution) (Miltenyi Biotec, Italy); mouse antihuman CD28 (CD28.2, #555726, 2 μg mL−1), mouse anti-human CD3 (UCHT1, #555330, 2 μg mL−1), goat anti-mouse (GAM, # 553998, 2 μg mL−1), anti-human CD3-PE (#555333, 1:10 dilution) (BD Biosciences, Milano, Italy); anti-human HLA-DR-PE (#12-9956-71, 1:10 dilution) (ThermoFisher Scientific, Rome, Italy).

Article Title: CD40- and 41BB-specific antibody fusion proteins with PDL1 blockade-restricted agonism
Article Snippet: After 4 days of coculture with ES-2scFv:CD3-mcherry cells, CD3 + T-cells were furthermore collected and stained for CD4 (anti-CD4-FITC; Immunotools), CD8 (anti-CD8-Pacific Blue; Beckman Coulter), CD25 (mouse anti-CD25-APC; Immunotools), mouse anti-41BB-APC (Immunotools) and with the zombie NIR fixable viability kit (Biolegend).



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