T cell co-culture was performed as previously described (78 (link)) with minor modifications. Briefly, tissue-culture plates were coated with 5 μg/mL purified anti-CD3 (clone HIT3a, Biolegend) at 4°C overnight and subsequently washed twice with 1X PBS. PBMCs from a healthy donor (Research Blood Components) were labelled with CFSE (Invitrogen) following the manufacturer’s protocol. PBMCs were resuspended in SFEM II (StemCell Technologies) with 5 μg/mL purified anti-CD28 (clone CD28.2, BioLegend) and plated at a density of 1M cells/mL. Isolated iMS1 or iMono cells were added at different ratios as indicated. The cells were left in culture for 3–4 days, with media replenished after 2 days. At the end of incubation, the cells were stained with CD3-AF700 (clone OKT3), CD4-APC (clone OKT4), and CD8a-PE (clone RPA-T8) (BioLegend) to determine the amount of CFSE dilution within the T cells.
Cd8a pe clone rpa t8
Manufactured by BioLegend
CD8a-PE (clone RPA-T8) is a monoclonal antibody that binds to the CD8 alpha chain. It is conjugated with the fluorescent dye Phycoerythrin (PE).
Automatically generated - may contain errors
Lab products found in correlation
Cd4 apc clone okt4, by BioLegend (2 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by BioLegend (2 mentions)
Cd3 af700 clone okt3, by BioLegend (2 mentions)
Sfemii, by STEMCELL (2 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Anti cd3 clone hit3a, by BioLegend (1 mentions)
Anti cd28 clone cd28, by BioLegend (1 mentions)
Clone hit3a, by BioLegend (1 mentions)
Carboxyfluorescein diacetate succinimidyl ester cfse, by Thermo Fisher Scientific (1 mentions)
Clone cd28, by BioLegend (1 mentions)
2 protocols using cd8a pe clone rpa t8
1
T Cell Co-Culture with Immune Cells
2
T Cell Proliferation Assay with iMS1 and iMono
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T cell coculture was performed as previously described (78 (link)) with minor modifications. Briefly, tissue culture plates were coated with purified anti-CD3 (5 μg/ml) (clone HIT3a, BioLegend) at 4°C overnight and subsequently washed twice with 1X PBS. PBMCs from a healthy donor (Research Blood Components) were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) (Invitrogen) following the manufacturer’s protocol. PBMCs were resuspended in SFEM II (StemCell Technologies) with purified anti-CD28 (5 μg/ml) (clone CD28.2, BioLegend) and plated at a density of 1 million cells/ml. Isolated iMS1 or iMono cells were added at different ratios as indicated. The cells were left in culture for 3 to 4 days, with media replenished after 2 days. At the end of incubation, the cells were stained with CD3-AF700 (clone OKT3), CD4-APC (clone OKT4), and CD8a-PE (clone RPA-T8) (BioLegend) to determine the amount of CFSE dilution within the T cells.
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