PBMC were analyzed using a 3-laser BD LSRII flow cytometer and FlowJo software (Treestar) as described (9 (link)). Primary antibodies were anti-Apc1 (Novus; NBP1-77375), anti-PGK1 (Abcam; ab67335), anti-γH2AX (Cell Signaling; #9718), and anti-phospho-NF-κB p65 (Ser536, P-p65) (Cell Signaling; #3033). Secondary antibodies were FITC goat anti-mouse Ig and FITC goat anti-rabbit Ig (BD Biosciences). Cell surface markers (BD Biosciences) were Pacific Blue mouse anti-human CD4, Alexa Fluor 700 mouse anti-human CD8, APC mouse anti-human CD19, and PE mouse anti-human CD14. Mouse anti-human HLA-DR was from BD Pharmingen (G46-6). Cyclosporin A (30024), Bay 11-7085 (B5681), and BI-78D3 (B3063) were from Sigma-Aldrich.
Alexa fluor 700 mouse anti human cd8
Manufactured by BD
Alexa Fluor 700 mouse anti-human CD8 is a fluorescently labeled antibody that recognizes the CD8 surface marker on human cells. It can be used for the detection and analysis of CD8-positive cells in flow cytometry applications.
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Lab products found in correlation
Apc mouse anti human cd19, by BD (1 mentions)
Fitc goat anti mouse ig, by BD (1 mentions)
Pe mouse anti human cd14, by BD (1 mentions)
Anti pgk1, by Abcam (1 mentions)
Mouse anti human hla dr, by BD (1 mentions)
Bi 78d3, by Merck Group (1 mentions)
Anti phospho nf κb p65 ser536 p p65, by Cell Signaling Technology (1 mentions)
Anti γh2ax, by Cell Signaling Technology (1 mentions)
Facsaria 2, by BD (1 mentions)
Flex t hla a 02 01 monomer uvx, by BioLegend (1 mentions)
Apc cy7 mouse anti human cd3, by BD (1 mentions)
2 protocols using alexa fluor 700 mouse anti human cd8
1
Flow Cytometric Analysis of PBMCs
2
Multi-color FACS Analysis and Sorting
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Cells were collected, washed, and resuspended in FACS buffer, and stained with fluorescent dye conjugated antibodies for 15 minutes at 4 °C. Peptide-MHC tetramers were generated through ultaviolet-irradiation-mediated peptide exchange method31 (link). APC-Cy™7 Mouse Anti-Human CD3 (BD Biosciences, 557832, 2.5 μl/5 × 105 cells), Alexa Fluor® 700 Mouse Anti-Human CD8 (BD Biosciences, 557945, 2.5 μl/5 × 105 cells), and Flex-T™ HLA-A*02:01 Monomer UVX (BioLegend, Cat#280004, 0.05 μg/1 × 106 cells) were used. After washing twice in FACS buffer, cells were analyzed using a FACSAria II (BD Biosciences) with live cell gating based on 4’,6-diamidino-2-phenylindole (DAPI) exclusion, and CD8+ pMHC-tetramer+ cells were sorted. The data were analyzed using FlowJo software (Tree Star).
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