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elispot colour development solution  (Bio-Rad)


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    Bio-Rad elispot colour development solution
    IFN-γ <t>ELISpot</t> assay showing the number of spot forming splenocytes in mice administered with either solar inactivated S . Typhimurium (green), <t>or</t> <t>PBS</t> (blue). LPS, FliC and BACT, were used as stimulants. Each data point (circle or square) represents an individual mouse, (from 1 experiment (Day 42)) and the error bars denote the interquartile range of the median. Data was analysed with a 2-way analysis of variance and post-hoc positive test, p values indicate significant differences between treatments groups for individual antigens.
    Elispot Colour Development Solution, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1040 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/elispot+substrate+solution/AP+Conjugate+Substrate+Kit/pmc07855330-113-16-21
    Average 99 stars, based on 1040 article reviews
    elispot colour development solution - by Bioz Stars, 2026-11
    99/100 stars

    Images

    1) Product Images from "Solar inactivated Salmonella Typhimurium induces an immune response in BALB/c mice"

    Article Title: Solar inactivated Salmonella Typhimurium induces an immune response in BALB/c mice

    Journal: Heliyon

    doi: 10.1016/j.heliyon.2021.e05903

    IFN-γ ELISpot assay showing the number of spot forming splenocytes in mice administered with either solar inactivated S . Typhimurium (green), or PBS (blue). LPS, FliC and BACT, were used as stimulants. Each data point (circle or square) represents an individual mouse, (from 1 experiment (Day 42)) and the error bars denote the interquartile range of the median. Data was analysed with a 2-way analysis of variance and post-hoc positive test, p values indicate significant differences between treatments groups for individual antigens.
    Figure Legend Snippet: IFN-γ ELISpot assay showing the number of spot forming splenocytes in mice administered with either solar inactivated S . Typhimurium (green), or PBS (blue). LPS, FliC and BACT, were used as stimulants. Each data point (circle or square) represents an individual mouse, (from 1 experiment (Day 42)) and the error bars denote the interquartile range of the median. Data was analysed with a 2-way analysis of variance and post-hoc positive test, p values indicate significant differences between treatments groups for individual antigens.

    Techniques Used: Enzyme-linked Immunospot

    Related Articles

    Ex Vivo:

    Article Title: N-terminal pyroglutamylation of an HLA-A24-restricted immunodominant epitope enhances SARS-CoV-2-specific T-cell responses
    Article Snippet: .. An ex vivo IFN-γ ELISpot assay was performed using the following antibodies and reagents: anti-mouse IFN-γ mAb AN18, purified (Mabtech, Code: 3321-3-1000), anti-mouse IFN-γ mAb R4-6A2, biotinylated (Mabtech, Code: 3321-6-1000), streptavidin-ALP (Mabtech, Code:3310-8-1000), 25×AP Color Development Buffer (Bio-Rad), AP Conjugated Substrate Kit (Bio-Rad, Cat# 1706432) according to the manufacturer’s protocol. .. Briefly, after washing a MultiScreen 96-well plate with sterilized PBS and blocked with RPMI 1640 medium (Thermo Fisher Scientific, Cat# 11875101) containing 10% FBS, 2×10 5 splenocytes from HLA-A24 transgenic mice immunized with CFA alone, the QI9/A24 peptide, and the pyrQI9 peptide 3 weeks after immunization per well were pulsed with the QI9/A24 or pyrQI9 peptide and cultured for 20 h. The responses were defined as the number of spot-forming cells (SFC)/2×10 5 calculated by ImmunoSpot (Cellular Technology Limited).

    Enzyme-linked Immunospot:

    Article Title: N-terminal pyroglutamylation of an HLA-A24-restricted immunodominant epitope enhances SARS-CoV-2-specific T-cell responses
    Article Snippet: .. An ex vivo IFN-γ ELISpot assay was performed using the following antibodies and reagents: anti-mouse IFN-γ mAb AN18, purified (Mabtech, Code: 3321-3-1000), anti-mouse IFN-γ mAb R4-6A2, biotinylated (Mabtech, Code: 3321-6-1000), streptavidin-ALP (Mabtech, Code:3310-8-1000), 25×AP Color Development Buffer (Bio-Rad), AP Conjugated Substrate Kit (Bio-Rad, Cat# 1706432) according to the manufacturer’s protocol. .. Briefly, after washing a MultiScreen 96-well plate with sterilized PBS and blocked with RPMI 1640 medium (Thermo Fisher Scientific, Cat# 11875101) containing 10% FBS, 2×10 5 splenocytes from HLA-A24 transgenic mice immunized with CFA alone, the QI9/A24 peptide, and the pyrQI9 peptide 3 weeks after immunization per well were pulsed with the QI9/A24 or pyrQI9 peptide and cultured for 20 h. The responses were defined as the number of spot-forming cells (SFC)/2×10 5 calculated by ImmunoSpot (Cellular Technology Limited).

    Purification:

    Article Title: N-terminal pyroglutamylation of an HLA-A24-restricted immunodominant epitope enhances SARS-CoV-2-specific T-cell responses
    Article Snippet: .. An ex vivo IFN-γ ELISpot assay was performed using the following antibodies and reagents: anti-mouse IFN-γ mAb AN18, purified (Mabtech, Code: 3321-3-1000), anti-mouse IFN-γ mAb R4-6A2, biotinylated (Mabtech, Code: 3321-6-1000), streptavidin-ALP (Mabtech, Code:3310-8-1000), 25×AP Color Development Buffer (Bio-Rad), AP Conjugated Substrate Kit (Bio-Rad, Cat# 1706432) according to the manufacturer’s protocol. .. Briefly, after washing a MultiScreen 96-well plate with sterilized PBS and blocked with RPMI 1640 medium (Thermo Fisher Scientific, Cat# 11875101) containing 10% FBS, 2×10 5 splenocytes from HLA-A24 transgenic mice immunized with CFA alone, the QI9/A24 peptide, and the pyrQI9 peptide 3 weeks after immunization per well were pulsed with the QI9/A24 or pyrQI9 peptide and cultured for 20 h. The responses were defined as the number of spot-forming cells (SFC)/2×10 5 calculated by ImmunoSpot (Cellular Technology Limited).

    Glutamate Assay:

    Article Title: Metabotropic Glutamate Receptor 3 Expression During Liver Disease Progression: Association with Inflammation and Cell Viability in Hepatocellular Carcinoma.
    Article Snippet: .. Diethylnitrosamine (DEN), (N0756), glutamate assay kit (MAK330), Minimum Essential Medium Eagle (MEM) (M0643), F12 Ham Kaighn’s Modification (F12K) (N3520), LY354740 (L1045), glutamate (L-glutamic acid monosodium salt, G5889), propidium iodide (P4170) were purchased from Sigma-Aldrich, St. Louis, MO, USA; Dulbecco’s Modified Eagle Medium (DMEM) low glucose from Gibco (Waltham, MA, USA); polyclonal rabbit anti-mGluR3 antibody (#AGC-012), mGluR3 blocking peptide (BLP-GC012) from Alomone (Jerusalem, Israel); polyclonal rabbit anti-mGluR3 antibody (ab140741), biotinylated goat anti-rabbit IgG antibody (ab6720), peroxidase–conjugated donkey anti-rabbit IgG antibody (AB97061) from Abcam (Cambridge, UK); monoclonal mouse anti-cAMP antibody (MAB2146) from R&D (Minneapolis, MN, USA); monoclonal rabbit anti-GAPDH (14C10), monoclonal rabbit b-actin (D6A8) from Cell Signaling Technology (Danvers, MA, USA); forskolin (1099) from Tocris (Minneapolis, MN, USA); RevertAid First Strand cDNA Synthesis Kit, Maxima SYBR Green qPCR Master Mix from Thermo Scientific (Waltham, MA, USA); TRIzolTM reagent, Alexa Fluor-488 donkey anti-rabbit IgG (A-21206), Alexa Fluor-488 goat anti-mouse IgG (H + L) (A-11001) from Invitrogen (Waltham, MA, USA); Vectashield® (H1000), Vectastain Elite ABC-HRP kit (PK-6100), DAB Substrate Kit, Peroxidase (HRP), (3,3′-diaminobenzidine) (SK-4100) from Vector Laboratories (Newark, CA, USA); DirectzolTM Miniprep RNA kit from Zymo Research (R2050) (Irving, CA, USA); AP conjugate substrate kit (1706432), protein assay dye (5000006) from Bio-Rad (Hercules, CA, USA). ..

    Modification:

    Article Title: Metabotropic Glutamate Receptor 3 Expression During Liver Disease Progression: Association with Inflammation and Cell Viability in Hepatocellular Carcinoma.
    Article Snippet: .. Diethylnitrosamine (DEN), (N0756), glutamate assay kit (MAK330), Minimum Essential Medium Eagle (MEM) (M0643), F12 Ham Kaighn’s Modification (F12K) (N3520), LY354740 (L1045), glutamate (L-glutamic acid monosodium salt, G5889), propidium iodide (P4170) were purchased from Sigma-Aldrich, St. Louis, MO, USA; Dulbecco’s Modified Eagle Medium (DMEM) low glucose from Gibco (Waltham, MA, USA); polyclonal rabbit anti-mGluR3 antibody (#AGC-012), mGluR3 blocking peptide (BLP-GC012) from Alomone (Jerusalem, Israel); polyclonal rabbit anti-mGluR3 antibody (ab140741), biotinylated goat anti-rabbit IgG antibody (ab6720), peroxidase–conjugated donkey anti-rabbit IgG antibody (AB97061) from Abcam (Cambridge, UK); monoclonal mouse anti-cAMP antibody (MAB2146) from R&D (Minneapolis, MN, USA); monoclonal rabbit anti-GAPDH (14C10), monoclonal rabbit b-actin (D6A8) from Cell Signaling Technology (Danvers, MA, USA); forskolin (1099) from Tocris (Minneapolis, MN, USA); RevertAid First Strand cDNA Synthesis Kit, Maxima SYBR Green qPCR Master Mix from Thermo Scientific (Waltham, MA, USA); TRIzolTM reagent, Alexa Fluor-488 donkey anti-rabbit IgG (A-21206), Alexa Fluor-488 goat anti-mouse IgG (H + L) (A-11001) from Invitrogen (Waltham, MA, USA); Vectashield® (H1000), Vectastain Elite ABC-HRP kit (PK-6100), DAB Substrate Kit, Peroxidase (HRP), (3,3′-diaminobenzidine) (SK-4100) from Vector Laboratories (Newark, CA, USA); DirectzolTM Miniprep RNA kit from Zymo Research (R2050) (Irving, CA, USA); AP conjugate substrate kit (1706432), protein assay dye (5000006) from Bio-Rad (Hercules, CA, USA). ..

    Blocking Assay:

    Article Title: Metabotropic Glutamate Receptor 3 Expression During Liver Disease Progression: Association with Inflammation and Cell Viability in Hepatocellular Carcinoma.
    Article Snippet: .. Diethylnitrosamine (DEN), (N0756), glutamate assay kit (MAK330), Minimum Essential Medium Eagle (MEM) (M0643), F12 Ham Kaighn’s Modification (F12K) (N3520), LY354740 (L1045), glutamate (L-glutamic acid monosodium salt, G5889), propidium iodide (P4170) were purchased from Sigma-Aldrich, St. Louis, MO, USA; Dulbecco’s Modified Eagle Medium (DMEM) low glucose from Gibco (Waltham, MA, USA); polyclonal rabbit anti-mGluR3 antibody (#AGC-012), mGluR3 blocking peptide (BLP-GC012) from Alomone (Jerusalem, Israel); polyclonal rabbit anti-mGluR3 antibody (ab140741), biotinylated goat anti-rabbit IgG antibody (ab6720), peroxidase–conjugated donkey anti-rabbit IgG antibody (AB97061) from Abcam (Cambridge, UK); monoclonal mouse anti-cAMP antibody (MAB2146) from R&D (Minneapolis, MN, USA); monoclonal rabbit anti-GAPDH (14C10), monoclonal rabbit b-actin (D6A8) from Cell Signaling Technology (Danvers, MA, USA); forskolin (1099) from Tocris (Minneapolis, MN, USA); RevertAid First Strand cDNA Synthesis Kit, Maxima SYBR Green qPCR Master Mix from Thermo Scientific (Waltham, MA, USA); TRIzolTM reagent, Alexa Fluor-488 donkey anti-rabbit IgG (A-21206), Alexa Fluor-488 goat anti-mouse IgG (H + L) (A-11001) from Invitrogen (Waltham, MA, USA); Vectashield® (H1000), Vectastain Elite ABC-HRP kit (PK-6100), DAB Substrate Kit, Peroxidase (HRP), (3,3′-diaminobenzidine) (SK-4100) from Vector Laboratories (Newark, CA, USA); DirectzolTM Miniprep RNA kit from Zymo Research (R2050) (Irving, CA, USA); AP conjugate substrate kit (1706432), protein assay dye (5000006) from Bio-Rad (Hercules, CA, USA). ..

    cDNA Synthesis:

    Article Title: Metabotropic Glutamate Receptor 3 Expression During Liver Disease Progression: Association with Inflammation and Cell Viability in Hepatocellular Carcinoma.
    Article Snippet: .. Diethylnitrosamine (DEN), (N0756), glutamate assay kit (MAK330), Minimum Essential Medium Eagle (MEM) (M0643), F12 Ham Kaighn’s Modification (F12K) (N3520), LY354740 (L1045), glutamate (L-glutamic acid monosodium salt, G5889), propidium iodide (P4170) were purchased from Sigma-Aldrich, St. Louis, MO, USA; Dulbecco’s Modified Eagle Medium (DMEM) low glucose from Gibco (Waltham, MA, USA); polyclonal rabbit anti-mGluR3 antibody (#AGC-012), mGluR3 blocking peptide (BLP-GC012) from Alomone (Jerusalem, Israel); polyclonal rabbit anti-mGluR3 antibody (ab140741), biotinylated goat anti-rabbit IgG antibody (ab6720), peroxidase–conjugated donkey anti-rabbit IgG antibody (AB97061) from Abcam (Cambridge, UK); monoclonal mouse anti-cAMP antibody (MAB2146) from R&D (Minneapolis, MN, USA); monoclonal rabbit anti-GAPDH (14C10), monoclonal rabbit b-actin (D6A8) from Cell Signaling Technology (Danvers, MA, USA); forskolin (1099) from Tocris (Minneapolis, MN, USA); RevertAid First Strand cDNA Synthesis Kit, Maxima SYBR Green qPCR Master Mix from Thermo Scientific (Waltham, MA, USA); TRIzolTM reagent, Alexa Fluor-488 donkey anti-rabbit IgG (A-21206), Alexa Fluor-488 goat anti-mouse IgG (H + L) (A-11001) from Invitrogen (Waltham, MA, USA); Vectashield® (H1000), Vectastain Elite ABC-HRP kit (PK-6100), DAB Substrate Kit, Peroxidase (HRP), (3,3′-diaminobenzidine) (SK-4100) from Vector Laboratories (Newark, CA, USA); DirectzolTM Miniprep RNA kit from Zymo Research (R2050) (Irving, CA, USA); AP conjugate substrate kit (1706432), protein assay dye (5000006) from Bio-Rad (Hercules, CA, USA). ..

    SYBR Green Assay:

    Article Title: Metabotropic Glutamate Receptor 3 Expression During Liver Disease Progression: Association with Inflammation and Cell Viability in Hepatocellular Carcinoma.
    Article Snippet: .. Diethylnitrosamine (DEN), (N0756), glutamate assay kit (MAK330), Minimum Essential Medium Eagle (MEM) (M0643), F12 Ham Kaighn’s Modification (F12K) (N3520), LY354740 (L1045), glutamate (L-glutamic acid monosodium salt, G5889), propidium iodide (P4170) were purchased from Sigma-Aldrich, St. Louis, MO, USA; Dulbecco’s Modified Eagle Medium (DMEM) low glucose from Gibco (Waltham, MA, USA); polyclonal rabbit anti-mGluR3 antibody (#AGC-012), mGluR3 blocking peptide (BLP-GC012) from Alomone (Jerusalem, Israel); polyclonal rabbit anti-mGluR3 antibody (ab140741), biotinylated goat anti-rabbit IgG antibody (ab6720), peroxidase–conjugated donkey anti-rabbit IgG antibody (AB97061) from Abcam (Cambridge, UK); monoclonal mouse anti-cAMP antibody (MAB2146) from R&D (Minneapolis, MN, USA); monoclonal rabbit anti-GAPDH (14C10), monoclonal rabbit b-actin (D6A8) from Cell Signaling Technology (Danvers, MA, USA); forskolin (1099) from Tocris (Minneapolis, MN, USA); RevertAid First Strand cDNA Synthesis Kit, Maxima SYBR Green qPCR Master Mix from Thermo Scientific (Waltham, MA, USA); TRIzolTM reagent, Alexa Fluor-488 donkey anti-rabbit IgG (A-21206), Alexa Fluor-488 goat anti-mouse IgG (H + L) (A-11001) from Invitrogen (Waltham, MA, USA); Vectashield® (H1000), Vectastain Elite ABC-HRP kit (PK-6100), DAB Substrate Kit, Peroxidase (HRP), (3,3′-diaminobenzidine) (SK-4100) from Vector Laboratories (Newark, CA, USA); DirectzolTM Miniprep RNA kit from Zymo Research (R2050) (Irving, CA, USA); AP conjugate substrate kit (1706432), protein assay dye (5000006) from Bio-Rad (Hercules, CA, USA). ..

    Real-time Polymerase Chain Reaction:

    Article Title: Metabotropic Glutamate Receptor 3 Expression During Liver Disease Progression: Association with Inflammation and Cell Viability in Hepatocellular Carcinoma.
    Article Snippet: .. Diethylnitrosamine (DEN), (N0756), glutamate assay kit (MAK330), Minimum Essential Medium Eagle (MEM) (M0643), F12 Ham Kaighn’s Modification (F12K) (N3520), LY354740 (L1045), glutamate (L-glutamic acid monosodium salt, G5889), propidium iodide (P4170) were purchased from Sigma-Aldrich, St. Louis, MO, USA; Dulbecco’s Modified Eagle Medium (DMEM) low glucose from Gibco (Waltham, MA, USA); polyclonal rabbit anti-mGluR3 antibody (#AGC-012), mGluR3 blocking peptide (BLP-GC012) from Alomone (Jerusalem, Israel); polyclonal rabbit anti-mGluR3 antibody (ab140741), biotinylated goat anti-rabbit IgG antibody (ab6720), peroxidase–conjugated donkey anti-rabbit IgG antibody (AB97061) from Abcam (Cambridge, UK); monoclonal mouse anti-cAMP antibody (MAB2146) from R&D (Minneapolis, MN, USA); monoclonal rabbit anti-GAPDH (14C10), monoclonal rabbit b-actin (D6A8) from Cell Signaling Technology (Danvers, MA, USA); forskolin (1099) from Tocris (Minneapolis, MN, USA); RevertAid First Strand cDNA Synthesis Kit, Maxima SYBR Green qPCR Master Mix from Thermo Scientific (Waltham, MA, USA); TRIzolTM reagent, Alexa Fluor-488 donkey anti-rabbit IgG (A-21206), Alexa Fluor-488 goat anti-mouse IgG (H + L) (A-11001) from Invitrogen (Waltham, MA, USA); Vectashield® (H1000), Vectastain Elite ABC-HRP kit (PK-6100), DAB Substrate Kit, Peroxidase (HRP), (3,3′-diaminobenzidine) (SK-4100) from Vector Laboratories (Newark, CA, USA); DirectzolTM Miniprep RNA kit from Zymo Research (R2050) (Irving, CA, USA); AP conjugate substrate kit (1706432), protein assay dye (5000006) from Bio-Rad (Hercules, CA, USA). ..

    Incubation:

    Article Title: Pathogenesis and immune responses to Eurasian avian like H1N1 and influenza D virus in pigs
    Article Snippet: After 48 h incubation at 37 °C with 5% CO2, plates were washed with PBS-T and incubated for one hour at room temperature with biotinylated anti-pig IFN-γ (0.5 μg/mL; clone P2C11, BD Pharmingen) or anti-pig IL-2 (0.5 μg/mL; clone MT265, Mabtech). .. Plates were washed and incubated with streptavidin–alkaline phosphatase (1:1000; S291, Invitrogen) for one hours at room temperature, then developed with BCIP/NBT substrate (1706432, Bio-Rad) according to the manufacturer’s instructions. ..



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    a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with <t>ELISpot.</t> Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.
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    Image Search Results


    a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with ELISpot. Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Cationic crosslinked carbon dots-adjuvanted intranasal vaccine induces protective immunity against Omicron-included SARS-CoV-2 variants

    doi: 10.1038/s41467-023-38066-8

    Figure Lengend Snippet: a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with ELISpot. Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.

    Article Snippet: Following additional five washes, TMB ELISpot substrate solution (Mabtech, Sweden) was added for spot formation.

    Techniques: Variant Assay, Enzyme-linked Immunospot, Clinical Proteomics, Expressing, Staining

    IFN-γ ELISpot assay showing the number of spot forming splenocytes in mice administered with either solar inactivated S . Typhimurium (green), or PBS (blue). LPS, FliC and BACT, were used as stimulants. Each data point (circle or square) represents an individual mouse, (from 1 experiment (Day 42)) and the error bars denote the interquartile range of the median. Data was analysed with a 2-way analysis of variance and post-hoc positive test, p values indicate significant differences between treatments groups for individual antigens.

    Journal: Heliyon

    Article Title: Solar inactivated Salmonella Typhimurium induces an immune response in BALB/c mice

    doi: 10.1016/j.heliyon.2021.e05903

    Figure Lengend Snippet: IFN-γ ELISpot assay showing the number of spot forming splenocytes in mice administered with either solar inactivated S . Typhimurium (green), or PBS (blue). LPS, FliC and BACT, were used as stimulants. Each data point (circle or square) represents an individual mouse, (from 1 experiment (Day 42)) and the error bars denote the interquartile range of the median. Data was analysed with a 2-way analysis of variance and post-hoc positive test, p values indicate significant differences between treatments groups for individual antigens.

    Article Snippet: Following incubation for at least 1 h at room temperature and washing six times with PBS, ELISpot colour development solution (170-6432, Bio-Rad, CA) was added.

    Techniques: Enzyme-linked Immunospot