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Anti granzyme b v450

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Anti-Granzyme B-V450 is a fluorescently labeled antibody that binds to the Granzyme B protein. Granzyme B is a serine protease involved in the cytotoxic T cell-mediated apoptosis pathway. The V450 fluorescent dye is conjugated to the antibody, allowing for its detection and quantification in flow cytometry applications.

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Lab products found in correlation

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Spelling variants of this product

Granzyme B (39 citations) Anti-granzyme B (17 citations) Granzyme B-V450 (2 citations)

4 protocols using anti granzyme b v450

1

Polyfunctional T-cell Profiling by Flow Cytometry

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ICS flow cytometry was done as previously described85 . In brief, 106 PBMCs were pulsed with autologous peptide at 10 μM in the presence of co-stimulatory antibodies (anti-CD28 and anti-CD49D), anti-CD107a-FITC, monensin, and brefeldin A (all from BD Biosciences) for 6 hrs. The cells were then surface stained with LIVE/DEAD cell dye (Invitrogen), anti-CD3-Alexa 780 (eBioscience), and anti-CD8-PE (BD Biosciences). The cells were permeabilized and labeled with anti-IFN-γ-Alexa 700, anti-IL-2-APC, anti-TNFα-PECy7, and anti-Granzyme B-V450 (all from BD Biosciences). CD3 events greater than 100,000 were acquired on an LSR II (BD Immunocytometry Systems), and data were analyzed using FlowJo (version 9.6.4; TreeStar). Polyfunctionality analysis was performed using Boolean gating and SPICE & PESTLE (version 5.1; NIAID)86 .
Carlson J.M., Du V.Y., Pfeifer N., Bansal A., Tan V.Y., Power K., Brumme C.J., Kreimer A., DeZiel C.E., Fusi N., Schaefer M., Brockman M.A., Gilmour J., Price M.A., Kilembe W., Haubrich R., John M., Mallal S., Shapiro R., Frater J., Harrigan P.R., Ndung’u T., Allen S., Heckerman D., Sidney J., Allen T.M., Goulder P.J., Brumme Z.L., Hunter E, & Goepfert P.A. (2016). Impact of Pre-adapted HIV Transmission. Nature medicine, 22(6), 606-613.
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2

Multiparametric Flow Cytometry Profiling of Immune Cells

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The following antibodies were used for flow cytometric analysis: Anti-Perforin-Alexa fluor 488, anti-CD28-allophycocyanin (APC), anti-CD19-APC, anti-CD3-APC, anti-CD56-APC, anti-CD3-APC-cyanin7 (Cy7), anti-CD8-APC-Cy7, anti-CD14-APC-Cy7, anti-CD14-fluorescein isothiocyanate (FITC), anti-HLA-DR-FITC, anti-CD16-R-phycoerythrin (PE), anti-GATA3-PE, anti-CD45RA-PE-cyanin 5 (Cy5), anti-CD4-PE-Cy5, anti-CD16-PE-Cy5, anti-CCR7-PE-Cy7, anti-CD4-PE-Cy7, anti-CD4-V450, anti-GranzymeB-V450, anti-CD8-V500, anti-CD3-V500 (all from BD Bioscience, Franklin Lakes, NJ), anti-CD57-FITC, anti-CD4-FITC, anti-CD85j-PE, anti-CX3CR1-PE, anti-T-bet-PE-Cy7, anti-Eomes-Peridinin chlorophyll (PerCP)-efluor710 (six from eBioscience, San Diego, CA), anti-HLA-DR-PE-Cy5, anti-IL-7Rα-V450, anti-CD57-V450 (three from BioLegend, San Diego, CA), anti-CX3CR1-FITC (MBL International Corporation, Woburn, MA). For intracellular staining of T cell lineage-specific transcription factors (T-bet, Gata3, and Eomes), granzyme B and perforin, PBMC were fixed and permeabilized with Fix/Perm buffer set (BioLegend). Stained cells were acquired by a BD LSRFortessa (BD bioscience) and analyzed by using FlowJo software (ver. 9.0 or 10.0; Tree Star, OR).
Kim H.Y., Yoo T.H., Hwang Y., Lee G.H., Kim B., Jang J., Yu H.T., Kim M.C., Cho J.Y., Lee C.J., Kim H.C., Park S, & Lee W.W. (2017). Indoxyl sulfate (IS)-mediated immune dysfunction provokes endothelial damage in patients with end-stage renal disease (ESRD). Scientific Reports, 7, 3057.
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3

Intracellular Cytokine and Effector Profiling

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Intracellular cytokine and effector molecule production was assessed using flow cytometry as described previously (40 (link)). Briefly, 106 cells were pulsed with peptide at a concentration of 10 μM in the presence of co-stimulatory antibodies (anti-CD28 and anti-CD49d), anti-CD107a-FITC, monensin, and brefeldin A (all from BD Biosciences). After 6 h, the cells were stained with a LIVE/DEAD dye (Invitrogen), anti-CD3-Alexa 780 (eBioscience), and anti-CD8-PE (BD Biosciences). The cells were then permeabilized and labeled with anti-IFN-γ-Alexa 700, anti-IL-2-allophycocyanin, anti-TNFα-PECy7, and anti-Granzyme B-V450 (all from BD Biosciences). At least 100,000 CD3+ events were acquired on an LSR II flow cytometer (BD Immunocytometry Systems), and data were analyzed using FlowJo software (version 9.6.4; TreeStar Inc.). Polyfunctionality analysis was performed using Boolean gating with SPICE & PESTLE software (version 5.1; NIAID).
Du V.Y., Bansal A., Carlson J., Salazar-Gonzalez J.F., Salazar M.G., Ladell K., Gras S., Josephs T.M., Heath S., Price D.A., Rossjohn J., Hunter E, & Goepfert P.A. (2016). HIV-1-specific CD8 T cells exhibit limited cross-reactivity during acute infection. Journal of immunology (Baltimore, Md. : 1950), 196(8), 3276-3286.
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4

Polyfunctional T-cell Profiling by Flow Cytometry

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ICS flow cytometry was done as previously described85 . In brief, 106 PBMCs were pulsed with autologous peptide at 10 μM in the presence of co-stimulatory antibodies (anti-CD28 and anti-CD49D), anti-CD107a-FITC, monensin, and brefeldin A (all from BD Biosciences) for 6 hrs. The cells were then surface stained with LIVE/DEAD cell dye (Invitrogen), anti-CD3-Alexa 780 (eBioscience), and anti-CD8-PE (BD Biosciences). The cells were permeabilized and labeled with anti-IFN-γ-Alexa 700, anti-IL-2-APC, anti-TNFα-PECy7, and anti-Granzyme B-V450 (all from BD Biosciences). CD3 events greater than 100,000 were acquired on an LSR II (BD Immunocytometry Systems), and data were analyzed using FlowJo (version 9.6.4; TreeStar). Polyfunctionality analysis was performed using Boolean gating and SPICE & PESTLE (version 5.1; NIAID)86 .
Carlson J.M., Du V.Y., Pfeifer N., Bansal A., Tan V.Y., Power K., Brumme C.J., Kreimer A., DeZiel C.E., Fusi N., Schaefer M., Brockman M.A., Gilmour J., Price M.A., Kilembe W., Haubrich R., John M., Mallal S., Shapiro R., Frater J., Harrigan P.R., Ndung’u T., Allen S., Heckerman D., Sidney J., Allen T.M., Goulder P.J., Brumme Z.L., Hunter E, & Goepfert P.A. (2016). Impact of Pre-adapted HIV Transmission. Nature medicine, 22(6), 606-613.
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