Flow cytometry was performed as described previously (6 (link)). Isolated cells were incubated with predetermined optimized concentrations of anti-CD3-Alexa fluor 700 (clone SP34-2, BD Pharmingen), anti-CD8-Pacific Blue or PE-CF594 (clone RPA-T8, BD Pharmingen), anti-CD20-APC-Cy7 (clone L27, BD Pharmingen), anti-CXCR5-phycoerythrin (PE) (Clone MU5UBEE, eBioscience), anti-CD200-APC (clone OX104, eBioscience), anti-CD4-PerCP (clone L200, BD Pharmingen), anti-CD95-PE-Cy5 (clone DX2, BD Pharmingen), anti-CCR7-APC-Cy7 (clone 3D12, Biolegend), anti-PD-1-PE-Cy7 (clone EH12.2H7, Biolegend), anti-ICOS-Pacific Blue (C398.4A, Biolegend) and Live/Dead dye-Alexa430 (Invitrogen). After staining the cell surface, cells were permeabilized and fixed by BD perm wash (BD Pharmingen) and washed twice. Finally, cells were incubated with anti-Ki67-FITC (B56, BD Pharmingen), washed twice and diluted in 1% PFA. Data were acquired on a LSRII flow cytometer (BD Biosciences) and the data analyzed using FlowJo software (version 9.2 Tree Star).
Anti cd95 pe cy5 clone dx2
Manufactured by BD
The Anti-CD95-PE-Cy5 (clone DX2) is a fluorescent-labeled antibody that binds to the CD95 (Fas) antigen. CD95 is a cell surface receptor that plays a key role in the regulation of programmed cell death or apoptosis. This antibody is conjugated with the fluorescent dyes PE-Cy5, allowing for the detection and analysis of CD95-expressing cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Perm wash, by BD (1 mentions)
Anti cd3 alexa fluor 700 clone sp34.2, by BD (1 mentions)
Anti cd4 percp clone l200, by BD (1 mentions)
Anti pd 1 pe cy7, by BioLegend (1 mentions)
Clone eh12.2h7, by BioLegend (1 mentions)
Anti ki67 fitc b56, by BD (1 mentions)
Anti cd20 apc cy7 clone l27, by BD (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Anti pd 1 bv421 clone eh12.2h7, by BioLegend (1 mentions)
Facs ariaιι, by BD (1 mentions)
Anti cd3 apc cy7 clone sp34 2, by BD (1 mentions)
2 protocols using anti cd95 pe cy5 clone dx2
1
Multiparameter Flow Cytometry Assay
2
Characterization of CD4+ Tfh Cells
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Mononuclear cells isolated from LN were stained with anti-CD3-APC-Cy7 (clone SP34-2, 5 μL, cat #557757), anti-CD28-PE-CF594 (clone CD28.2, 5 μL, cat #562296), and anti-CD95-PECy5 (clone DX2, 10 μL, cat #559773) from BD Biosciences; anti-CD4-BV650 (clone OKT4, 2 μL, cat #317436), anti-PD-1-BV421 (clone EH12.2H7, 5 μL, cat # 3299220), and anti-CD200-PE (clone OX-104, 5 μL, cat #329206) from Biolegend; anti-CD8-FITC (clone 3B5, 5 μL, cat #MHCD0801-4) from Thermo Fisher Scientific; and Aqua Live/Dead Fixable Aqua from Invitrogen (AmCyan, 2 μL of 1:20 PBS dilution, cat. L34957). Sorting of CD4+ Tfh (PD1+CD200hi) was performed using a FACS AriaΙΙ (BD Biosciences) in samples collected before, during FTY720 treatment, at ART interruption, and 29 days after ART interruption. Post-sorting FACS analysis determined that sorted CD4+ T cell subsets were on average >96% pure.
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