Spleens from the indicated mice were collected and dissociated, followed by filtering through a cell strainer in PBS with 1% FBS. Cells were collected by centrifugation at 1500 rpm for 5 min, red blood cells were removed by red blood cell lysis buffer, and the remain cells were resuspended and incubated with antibodies (CD3-BV421, CD4-PECy7, CD8-BB515, CD69-BV711, Ly6c-BV786, CD44-APC, CD62L-PE, all from BD Biosciences) on ice in the dark (or wrap in foil) for 20 min. The strained cells were resuspended in PBS and analyzed with FACSymphony™ A5 instrument (BD Biosciences).
Cd8 bb515
Manufactured by BD
The CD8-BB515 is a fluorescently labeled antibody that binds to the CD8 protein, which is expressed on the surface of certain T cells. It can be used to identify and quantify CD8+ T cells in various biological samples through flow cytometry or other immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Cd69 bv711, by BD (1 mentions)
Cd44 apc, by BD (1 mentions)
Cd62l pe, by BD (1 mentions)
Cd3 bv421, by BD (1 mentions)
Cd4 pe cy7, by BD (1 mentions)
Facsymphony a5, by BD (1 mentions)
Cd16 apc, by BD (1 mentions)
Cd14 bv510, by BD (1 mentions)
Cd56 buv395, by BD (1 mentions)
Cd4 pe, by BD (1 mentions)
Lsrfortessa x 20 flow cytometer, by BD (1 mentions)
Cd3 pe cy7, by BD (1 mentions)
Protein transport inhibitor, by BD (1 mentions)
Cd107a bv421, by BD (1 mentions)
Cd4 bb700, by BD (1 mentions)
Cd3 apc cy7, by BioLegend (1 mentions)
3 protocols using cd8 bb515
1
Murine Splenocyte Phenotyping
2
Multicolor Flow Cytometry Panel for Leukocyte Profiling
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To evaluate the leukocyte populations, 5 × 105 cells of the thawed PBMCs were seeded in 96-well plates (Additional file 1: Figure S1 ) and stained with a panel of humans antibodies CD4-PE (Clone, RPA-T4 BD bioscience), CD8-BB515 (Clone, RPA-T8 BD bioscience), CD3-PeCy7 (Clone, SK7 BD bioscience), CD14-BV510 (Clone, M0P9 BD bioscience), CD16-APC (Clone, 3G8 BD bioscience) and CD56-BUV-395 (Clone, NCAM16.2 BD bioscience), according to manufacturer instructions. Cells were resuspended in 100 ul of PBS 1%FBS containing the mix of antibodies and incubated at 4°C for 30 min. After incubation, cells were washed twice with PBS and centrifuged at 400 g for 5 min at 4°C. Then, the cell pellet was resuspended in 200 ul of PBS 1%FBS. Samples were acquired at the LSRFortessa X20 flow cytometer (BD Biosciences) and analyzed using FlowJo v10.0.7 software (BD Biosciences). Gating strategy used to identify lymphoid and myeloid subsets is showed in additional file 1: Figure S1 .
3
CAR-T Cell Activation Assay
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A 1: 1 ratio was used in co-culturing NT or CAR-T cells pretreated with Dec (0.05, 0.1μM) or Aza (0.5, 1μM) with MV4-11 cells for 16 h. Meanwhile CD107a-BV421 (BD Biosciences, Cat#562623) and protein transport inhibitor (BD Biosciences, Cat#554724) were added to the culture medium. After cells collection, Antibodies CD3-APC/Cy7 (Biolegend, Cat#300318), CD4-BB700 (BD Biosciences, Cat# 566392), and CD8-BB515 (BD Biosciences, Cat# 564526) were applied to stain the cells.
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