Anti cd8α pacific blue clone rpa t8

Manufactured by BD

Anti-CD8α Pacific Blue (clone RPA-T8) is a fluorochrome-conjugated monoclonal antibody used for the identification and enumeration of CD8+ T cells in flow cytometry applications.

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Lab products found in correlation

Anti cd3 alexa fluor 700 clone sp43 2, by BD (2 mentions) Anti cd4 allophycocyanin clone l200, by BD (2 mentions) Live dead amine reactive viability dye, by Thermo Fisher Scientific (2 mentions) Facs fortessa, by BD (1 mentions) Facsymphony s6 cell sorter, by BD (1 mentions) Lymphocyte separation media, by MP Biomedicals (1 mentions)

Close spelling variants of this product

RPA-T8 (53 citations) Anti-CD8α (15 citations) Clone RPA-T8 (14 citations) Anti-CD8α-Pacific Blue (3 citations) Pacific Blue–CD8α (2 citations)

2 protocols using anti cd8α pacific blue clone rpa t8

Comprehensive T Cell Phenotyping

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T cell phenotyping for all experiments was assessed with antibodies reactive to the following surface antigens: anti-CD3 Alexa Fluor 700 (clone SP43-2, catalog 557917), anti-CD4 Allophycocyanin (clone L200, catalog 551980), anti-CD8α Pacific Blue (clone RPA-T8, catalog 558207), and anti-CD69 Phycoerythrin.cy7 (PE-cy7) (clone FN50, catalog 557745) (all from BD). Cells were washed once in ice-cold PBS and incubated with fluorochrome-labeled antibodies at 4°C with the addition of LIVE/DEAD amine-reactive viability dye (Thermo Fisher Scientific). After 20 minutes, cells were washed a second time in PBS. For experiments requiring only strict phenotyping, cells were fixed with 1% PFA and acquired on a BD FACS Fortessa.

Mudd J.C., Lai S., Shah S., Rahmberg A., Flynn J.K., Starke C.E., Perkins M.R., Ransier A., Darko S., Douek D.C., Hirsch V.M., Cameron M, & Brenchley J.M. (2020). Epigenetic silencing of CD4 expression in nonpathogenic SIV infection in African green monkeys. JCI Insight, 5(18), e139043.

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Isolation and Characterization of Peripheral Blood Lymphocytes

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Cells were isolated from fresh peripheral blood by density gradient centrifugation using lymphocyte separation media (MP Biomedicals, #0850494-CF), washed in PBS, and counted. Approximately 5e6 cells were then resuspended in 100μl PBS. Cells were stained with LIVE/DEAD amine-reactive viability dye (Thermo Fisher Scientific, #L34966) and a panel of antibodies, including anti-CD3 alexa fluor 700 (clone SP43–2, BD Biosciences, #557917), anti-CD4 allophycocyanin (clone L200, BD, #551980), anti-CD8α pacific blue (clone RPA-T8, BD, #558207), CD28 energy-coupled dye [ECD] (clone 28.2, Beckman Coulter, Brea, CA, #6607111), CD95 Cy5-PE (clone DX2, BD, #561977), for 20 minutes at 4°C. Cells were washed in PBS and resuspended in complete RPMI with 10% FBS. For each population, 30,000 cells were sorted using a FACSymphony S6 cell sorter (BD).

Rahmberg A.R., Markowitz T.E., Mudd J.C., Hirsch V, & Brenchley J.M. (2022). Epigenetic reprogramming leads to down regulation of CD4 and functional changes in African green monkey memory CD4+ T cells. Journal of immunology (Baltimore, Md. : 1950), 209(2), 337-345.

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