Magnetic Cell Separation:Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were harvested, washed twice with auto MACS Rinsing Solution (Miltenyi), reacted with the following antibodies, and the fractions of immune cells present was analyzed by FACS analysis: reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: HI100), anti-CD25 antibody (BD, clone: 2A3), anti-CD16 antibody (Biolegend, clone: 3G8), anti-CD56 antibody (Biolegend, clone: HCD56), anti-CTLA4 antibody (Biolegend, clone: BNI3). .. FACS analysis was performed by BD LSR Fortessa X-20 (BD).
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were harvested, washed twice with auto MACS Rinsing Solution (Miltenyi), reacted with the following antibodies, and the fractions of immune cells present was analyzed by FACS analysis: reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: HI100), anti-CD25 antibody (BD, clone: 2A3), anti-CD16 antibody (Biolegend, clone: 3G8), anti-CD56 antibody (Biolegend, clone: HCD56), anti-CTLA4 antibody (Biolegend, clone: BNI3). .. FACS analysis was performed by BD LSR Fortessa X-20 (BD).
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were collected and washed twice with auto MACS Rinsing Solution (Milteny), and then allowed to react with the following antibodies to analyze the fraction of immune cells present with FACS analysis. .. Reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: 5 Hg), anti-CD25 antibody (BD, clone: 2A3), anti-CTLA4 antibody (Biolegend, clone: BNI3).
Article Title: Riboflavin photochemical therapy triggered PANoptosis by inducing mitochondrial damage in circulating colorectal cancer cells.
Article Snippet: The pelleted PBMCs were resuspended in complete RPMI-1640 medium supplemented with 10 % fetal bovine serum, 1 % penicillin/streptomycin, 0.1 % β-mercaptoethanol (Gibco), and 10 ng/mL recombinant human IL-2 (#I2644, Sigma). .. Whole Blood CD326 (EpCAM) MicroBeads, human (#130-109-828), Whole Blood Column Kit (#130-093-545), LS Separation Columns (#130-122-729), Auto MACS Rinsing Solution (#130-091-222), and MACS BSA Stock Solution (#130-091-376) was purchased from Miltenyi Biotec. .. Briefly, 50 μL of Straight From Whole Blood CD326 MicroBeads were added to whole blood, thoroughly mixed, and incubated for 30 min at 2–8 ◦C.
Article Title: Redefining Dendritic Cell Vaccines: Synergistically Co-priming DC and B Cells With Nanoparticles Loading Whole Cell Antigens Maximizes the Efficacy of DC Vaccines.
Article Snippet: PBS, DMEM medium, 1640 medium, fetal bovine serum, trypsin EDTA solution, and penicillin-streptomycin solution were purchased from Wuhan Pricella Biotechnology Co.,Ltd. .. Anti-PD-L1 antibody (STARTER, catalog number S0B0593, Clone: 10F.9G2), Pan B Cell Isolation Kit II, mouse (Miltenyi, catalog number 130-104-443), LD Columns (Miltenyi, catalog number 130-042-901-1), and auto MACS Running Buffer (Miltenyi, catalog number 130-091-221- 1) were obtained from Shanghai Universal Biotech Co., Ltd. IL-4 (PeproTech, catalog number 214-14, Source: E. coli), GMCSF (PeproTech, catalog number 315-03, Source: E. coli), M-CSF (PeproTech, catalog number 315-02, Source: E. coli), IL-15 (PeproTech, catalog number 210-15, Source: E. coli), TNF- α (PeproTech, catalog number 315-01A, Source: E. coli), IL-6 (PeproTech, catalog number 216-16, Source: E. coli), IL-1 β (PeproTech, catalog number 2111-11B, Source: E. coli), and IL-2 (PeproTech, catalog number 212-12, Source: E. coli) were obtained from Dakewe Biotech Co., Ltd. Zombie Aqua Fixable Viability Kit (BioLegend, catalog number 423102, dilution: 1:200), anti-mouse CD16/32 (BioLegend, catalog number 101320, clone: 93, dilution: 1:200), 14 of 20 APC/Cyanine7 anti-mouse B220 (BioLegend, catalog number 103224, clone: RA3-6B2, dilution: 1:200), PerCP/Cyanine5.5 antimouse CD11c (BioLegend, catalog number 117328, clone: N418, dilution: 1:200), PE/Cyanine7 anti-mouse CD86 (BioLegend, catalog number 159208, clone: A17199A, dilution: 1:200), FITC anti-mouse I-A/l-E (BioLegend, catalog number 107606, clone: M5/114.15.2, dilution: 1:200), APC anti-mouse CD80 (BioLegend, catalog number 104714, clone: 16-10A1, dilution: 1:200), PE anti-mouse-H-2 kb (BioLegend, catalog number 116507, clone: AF6-88.5, dilution: 1:200), APC anti-mouse CD40 (BioLegend, catalog number 124612, clone: 3/23, dilution: 1:200), PE antimouse CD154 (CD40L) (BioLegend, catalog number 158104, clone: QA17A53, dilution: 1:200), PE anti-mouse CD274 (B7H1, PD-L1) Antibody (BioLegend, catalog number 124308, clone: 10F.9G2, dilution: 1:200) were purchased from Dakewei Biotech Co., Ltd. ..
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After six hours, PBMCs were collected, washed with auto MACS Rinsing Solution (Milteny), and then reacted with the following antibodies to analyze existing immune cell fractions by FACS analysis: a reagent to determine viability (Biolegend, Zombie NIR), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD45RA antibody (Biolegend, clone: HI100), anti-FoxP3 antibody (eBioscience, clone: 236A/E7), and anti-CTLA4 antibody (BD, clone: BNI3). .. The FACS analysis was performed with BD FACSCelesta (BD).
FACS:Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were harvested, washed twice with auto MACS Rinsing Solution (Miltenyi), reacted with the following antibodies, and the fractions of immune cells present was analyzed by FACS analysis: reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: HI100), anti-CD25 antibody (BD, clone: 2A3), anti-CD16 antibody (Biolegend, clone: 3G8), anti-CD56 antibody (Biolegend, clone: HCD56), anti-CTLA4 antibody (Biolegend, clone: BNI3). .. FACS analysis was performed by BD LSR Fortessa X-20 (BD).
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were harvested, washed twice with auto MACS Rinsing Solution (Miltenyi), reacted with the following antibodies, and the fractions of immune cells present was analyzed by FACS analysis: reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: HI100), anti-CD25 antibody (BD, clone: 2A3), anti-CD16 antibody (Biolegend, clone: 3G8), anti-CD56 antibody (Biolegend, clone: HCD56), anti-CTLA4 antibody (Biolegend, clone: BNI3). .. FACS analysis was performed by BD LSR Fortessa X-20 (BD).
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were collected and washed twice with auto MACS Rinsing Solution (Milteny), and then allowed to react with the following antibodies to analyze the fraction of immune cells present with FACS analysis. .. Reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: 5 Hg), anti-CD25 antibody (BD, clone: 2A3), anti-CTLA4 antibody (Biolegend, clone: BNI3).
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After six hours, PBMCs were collected, washed with auto MACS Rinsing Solution (Milteny), and then reacted with the following antibodies to analyze existing immune cell fractions by FACS analysis: a reagent to determine viability (Biolegend, Zombie NIR), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD45RA antibody (Biolegend, clone: HI100), anti-FoxP3 antibody (eBioscience, clone: 236A/E7), and anti-CTLA4 antibody (BD, clone: BNI3). .. The FACS analysis was performed with BD FACSCelesta (BD).
Recombinase Polymerase Amplification:Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were harvested, washed twice with auto MACS Rinsing Solution (Miltenyi), reacted with the following antibodies, and the fractions of immune cells present was analyzed by FACS analysis: reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: HI100), anti-CD25 antibody (BD, clone: 2A3), anti-CD16 antibody (Biolegend, clone: 3G8), anti-CD56 antibody (Biolegend, clone: HCD56), anti-CTLA4 antibody (Biolegend, clone: BNI3). .. FACS analysis was performed by BD LSR Fortessa X-20 (BD).
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After 6 hours, PBMCs were harvested, washed twice with auto MACS Rinsing Solution (Miltenyi), reacted with the following antibodies, and the fractions of immune cells present was analyzed by FACS analysis: reagent for determining viability (Biolegend, Zombie Aqua), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD8 antibody (BD, clone: SK1), anti-CD45RA antibody (Biolegend, clone: HI100), anti-CD25 antibody (BD, clone: 2A3), anti-CD16 antibody (Biolegend, clone: 3G8), anti-CD56 antibody (Biolegend, clone: HCD56), anti-CTLA4 antibody (Biolegend, clone: BNI3). .. FACS analysis was performed by BD LSR Fortessa X-20 (BD).
Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: .. After six hours, PBMCs were collected, washed with auto MACS Rinsing Solution (Milteny), and then reacted with the following antibodies to analyze existing immune cell fractions by FACS analysis: a reagent to determine viability (Biolegend, Zombie NIR), anti-CD3 antibody (BD, clone: UCHT1), anti-CD4 antibody (BD, clone: RPA-T4), anti-CD45RA antibody (Biolegend, clone: HI100), anti-FoxP3 antibody (eBioscience, clone: 236A/E7), and anti-CTLA4 antibody (BD, clone: BNI3). .. The FACS analysis was performed with BD FACSCelesta (BD).
Blocking Assay:
other:Article Title: Anti-CTLA-4 antibody and use thereof
Article Snippet: Colon 38 cells were maintained and passaged in the RPMI1640 medium (NACALAI TESQUE, INC.) containing 10% FBS (CORNING). (8-1-2) Production of a Syngeneic Tumor Line Transplant Mouse Model Human CTLA4 KI, human CD3 EDG replaced mice (hCTLA4 KI hCD3 EDG replaced mice) (12-week-old, male, or 7-week-old, female), which are a hybrid strain of human CTLA4 knock-in mice (Blood (2005) 106(9): 3127-3133) and in-house-produced human CD3 EDG replaced mice (Sci Rep (2017) 7:45839), were used.
Cell Isolation:Article Title: Redefining Dendritic Cell Vaccines: Synergistically Co-priming DC and B Cells With Nanoparticles Loading Whole Cell Antigens Maximizes the Efficacy of DC Vaccines.
Article Snippet: PBS, DMEM medium, 1640 medium, fetal bovine serum, trypsin EDTA solution, and penicillin-streptomycin solution were purchased from Wuhan Pricella Biotechnology Co.,Ltd. .. Anti-PD-L1 antibody (STARTER, catalog number S0B0593, Clone: 10F.9G2), Pan B Cell Isolation Kit II, mouse (Miltenyi, catalog number 130-104-443), LD Columns (Miltenyi, catalog number 130-042-901-1), and auto MACS Running Buffer (Miltenyi, catalog number 130-091-221- 1) were obtained from Shanghai Universal Biotech Co., Ltd. IL-4 (PeproTech, catalog number 214-14, Source: E. coli), GMCSF (PeproTech, catalog number 315-03, Source: E. coli), M-CSF (PeproTech, catalog number 315-02, Source: E. coli), IL-15 (PeproTech, catalog number 210-15, Source: E. coli), TNF- α (PeproTech, catalog number 315-01A, Source: E. coli), IL-6 (PeproTech, catalog number 216-16, Source: E. coli), IL-1 β (PeproTech, catalog number 2111-11B, Source: E. coli), and IL-2 (PeproTech, catalog number 212-12, Source: E. coli) were obtained from Dakewe Biotech Co., Ltd. Zombie Aqua Fixable Viability Kit (BioLegend, catalog number 423102, dilution: 1:200), anti-mouse CD16/32 (BioLegend, catalog number 101320, clone: 93, dilution: 1:200), 14 of 20 APC/Cyanine7 anti-mouse B220 (BioLegend, catalog number 103224, clone: RA3-6B2, dilution: 1:200), PerCP/Cyanine5.5 antimouse CD11c (BioLegend, catalog number 117328, clone: N418, dilution: 1:200), PE/Cyanine7 anti-mouse CD86 (BioLegend, catalog number 159208, clone: A17199A, dilution: 1:200), FITC anti-mouse I-A/l-E (BioLegend, catalog number 107606, clone: M5/114.15.2, dilution: 1:200), APC anti-mouse CD80 (BioLegend, catalog number 104714, clone: 16-10A1, dilution: 1:200), PE anti-mouse-H-2 kb (BioLegend, catalog number 116507, clone: AF6-88.5, dilution: 1:200), APC anti-mouse CD40 (BioLegend, catalog number 124612, clone: 3/23, dilution: 1:200), PE antimouse CD154 (CD40L) (BioLegend, catalog number 158104, clone: QA17A53, dilution: 1:200), PE anti-mouse CD274 (B7H1, PD-L1) Antibody (BioLegend, catalog number 124308, clone: 10F.9G2, dilution: 1:200) were purchased from Dakewei Biotech Co., Ltd. ..
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