Fluorochrome-conjugated antibodies were from eBioscience or BioLegend: CD3 (OKT3), CD4 (OKT4), CD14 (63D3), CD11b (ICRF44), CD19 (HIB19), CD27 (LG.3A10), CD45RA (HI100), CD107a (H4A3), TCR γ/δ (B1), Vδ2 (B6), Vδ1 (TS8.2), Granzyme B (QA16A02), IFN-γ (B27), PD-1 (EH12.1), CTLA4 (BNI3), T-bet (4B10), Eomes (WD1928), CD80 (2D10) and CD86 (IT2.2). For surface markers, expanded Vγ9Vδ2 T cells or PBMCs were stained with diluted fluorochrome-conjugated antibodies on ice and washed with FACS buffer; and for intracellular biomarkers, cells were permeabilized and stained with antibodies diluted in permeabilization buffer (BioLegend, San Diego, CA). Samples were acquired using a LSR-A flow cytometer (BD Biosciences, Baltimore, MD) and analyzed using FlowJo software (Tree Star, Ashland, OR).
Lsr a flow cytometer
Manufactured by BD
The LSR-A flow cytometer is a laboratory instrument used for the analysis and sorting of cells or particles suspended in a fluid stream. It is designed to measure and analyze multiple physical and fluorescent characteristics of individual cells or particles as they pass through a laser beam.
Automatically generated - may contain errors
Lab products found in correlation
Permeabilization buffer, by BioLegend (1 mentions)
Mouse fc block, by BD (1 mentions)
Apc conjugated anti cd19 1d3, by BD (1 mentions)
Pe cy7 conjugated anti cd45 30 f11, by BD (1 mentions)
V450 conjugated anti cd11c hl3, by BD (1 mentions)
Pe conjugated anti f4 80 bm8, by Thermo Fisher Scientific (1 mentions)
Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (1 mentions)
2 protocols using lsr a flow cytometer
1
Multicolor Flow Cytometry Analysis
2
Detailed Multicolor Flow Cytometry Staining
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Single cells suspensions were blocked with Mouse BD Fc Block™ (BD Biosciences, Franklin Lakes, NJ) for 10 min and then stained in FACS buffer (PBS with 4% FBS and 2 mM EDTA) with the following fluorochrome-conjugated antibodies: APC-conjugated anti-CD19 (1D3, #550992, 1:200), APC-Cy7-conjugated anti-CD4 (GK1.5, #552051, 1:200), PE-Cy7-conjugated anti-CD8a (53-6.7, #552877, 1:200), FITC-conjugated anti-CD45 (30-F11, #561088, 1:100), V450-conjugated anti-CD3e (500A2, 560801, 1:100), APC-Cy7-conjugated anti-Ly6C (AL-21, #560596, 1:100), PE-Cy7-conjugated anti-CD45 (30-F11, #552848, 1:100), V450-conjugated anti-CD11c (HL3, #560521, 1:100), PerCP-Cy5.5-conjugated anti-CD11b (M1/70, #561114, 1:100), AF700-conjugated anti-Ly6G (1A8, #561236, 1:100) (from BD Biosciences); PE-conjugated anti-F4/80 (BM8, #12-4801, 1:100), AF700-conjugated anti-CD25 (PC61.5, #56-0251-80, 1:100) (from eBioscience, San Diego, CA). Viability was determined by staining cells with LIVE/DEAD™ Fixable Aqua Dead Cell stain, 1:300 (Thermo Fisher Scientific, Waltham, MA). Flow cytometry was performed on a LSR A flow cytometer (BD Biosciences), and data were analyzed using FlowJo software.
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