Spleen and LN cells were incubated with Fixable Viability Dye conjugated with eFluor780 fluorochrome (Affymetrix, eBioscience, Santa Clara, CA, USA), and background staining was blocked with anti‐FcγR mAb (clone 2.4G2). Cells were then incubated for 15 minutes at 4°C with H‐2k(d) AMQMLKETI APC‐labelled Tetramer (Tetr‐gag, NIH Tetramer Core Facility, Atlanta, GA) and PE‐labelled Pentamer (Pent‐gag, Proimmune, Oxford, UK) to stain for gag197‐205(gag)‐specific CD8 T cells. Cells were incubated for further 15 minutes at 4°C after addition of the following mAbs: anti‐CD3 peridinin chlorophyll protein (PerCP)‐Cy5.5 (clone 145‐2C11; BD Biosciences, San Jose, CA, USA), anti‐CD8α BUV805 (clone 53‐6.7, BD Biosciences) and anti‐CD62L phycoerythrin (PE)‐Cy7 (clone MEL‐14, Biolegend, San Diego, CA, USA). Blood samples were incubated for 30 minutes at RT with the above antibodies/reagents that were placed all together. After washing, blood cells were fixed with Cell Fix solution (BD Biosciences). Red cells were lysed with Pharm Lyse solution (BD Biosciences).
Anti cd8α buv805 clone 53 6
Manufactured by BD
Sourced in United States
Anti-CD8α BUV805 (clone 53-6.7) is a fluorochrome-conjugated monoclonal antibody that binds to the CD8α molecule. CD8α is a transmembrane glycoprotein expressed on the surface of cytotoxic T cells and a subset of natural killer cells. The BUV805 fluorochrome allows detection of the antibody-antigen interaction using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Pharm lyse solution, by BD (1 mentions)
Fixable viability dye conjugated with efluor780 fluorochrome, by Thermo Fisher Scientific (1 mentions)
Cell fix solution, by BD (1 mentions)
Anti cd62l pecy7, by BioLegend (1 mentions)
Anti cd127 biotin clone a7r34, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 700, by Thermo Fisher Scientific (1 mentions)
Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions)
Ebioscience fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions)
Streptavidin pe cy7, by BD (1 mentions)
2 protocols using anti cd8α buv805 clone 53 6
1
Quantification of Gag-specific CD8+ T Cells
2
Comprehensive Multiparameter Analysis of Antigen-Specific T Cells
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Cells were incubated with purified anti-mouse CD16/CD32 clone 2.4G2 (Fc block; BD Biosciences, San Jose, CA, USA), and stained as described with H-2k(d) AMQMLKETI (gag197-205) allophycocyanin (APC)-labeled Tetramer (Tetr-gag, NIH Tetramer Core Facility, Atlanta, GA, USA) and phycoerythrin (PE)-labeled Pentamer (Pent-gag, Proimmune, Oxford, UK), fluorochrome conjugated monoclonal Antibodies (mAbs) against surface (CD3, CD8, CD127, CD62L) and intracellular (Ki-67) molecules, and Hoechst 33342 (Thermo Fisher Scientific, Waltham, MA, USA) (14 (link)). The following mAbs were used: anti-CD3ε peridinin chlorophyll protein (PerCP)-Cy5.5 (clone 145-2C11, BD Biosciences), anti-CD8α BUV805 (clone 53-6.7, BD Biosciences), anti-CD127 biotin (clone A7R34, eBioscience, Thermo Fisher Scientific) plus Streptavidin PE-Cy7 (BD Biosciences), or anti-CD62L PE-Cy7 (clone MEL-14, Biolegend, San Diego, CA, USA), and anti-Ki-67 mAb conjugated with Fluorescein isothiocyanate (FITC) or Alexafluor 700 (clone SolA-15; eBioscience, Thermo Fisher Scientific). Dead cells were excluded with eBioscience Fixable Viability Dye eFluor780 (eFluor780, Invitrogen, Thermo Fisher Scientific)
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