PBMC were thawed at 37°C, transferred into pre-warmed complete RPMI10 culture medium, pelleted by centrifugation, resuspended in RPMI10, and left to rest at 37˚C for 1.5 h. Cells were stained with a cocktail of cell surface antibodies in FACS buffer (PBS + 2% FCS) for 30min at room temperature (RT) in the dark. Cells were washed and resuspended in FACS buffer, 7AAD viability dye (Biolegend) was added, and cells were acquired on a BD FACSymphony A5 flow cytometer (BD Biosciences). Data were analysed by FlowJo V10 (BD Biosciences). The following anti-human flow cytometry antibodies were used: TCR-gd-FITC (B1, Biolegend), CD19-PerCP-Cy5.5 (HIB19, Biolegend), CD14-PE (M5E2, Biolegend), CD56-PE-CF594 (NCAM16.2, BD Biosciences), CCR7-PE-Cy7 (G043H7, Biolegend), PD-1-AF700 (EH12.2H7, Biolegend), CD45RO-APC-Cy7 (UCHL1, BD Biosciences), CD127-BV421 (A019D5, Biolegend), CD28-BV480 (CD28.2, BD Biosciences), CD20-BV605 (L27, BD Biosciences), CD16-BV650 (3G8, BD Biosciences), HLA-DR-BV711 (L243, Biolegend), CD4-BV786 (SK3, Biolegend), CD3-BUV395 (UCHT1, BD Biosciences), CD8-BUV496 (RPA-T8, BD Biosciences), CD25-BUV737 (2A3, BD Biosciences).
Tcr gd fitc b1
Manufactured by BioLegend
TCR-gd-FITC (B1) is a fluorescent-labeled monoclonal antibody that binds to the gamma-delta T cell receptor (TCR) on human cells. This antibody is conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate) and can be used to detect and analyze gamma-delta T cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Cd20 bv605, by BD (1 mentions)
7 aad viability dye, by BioLegend (1 mentions)
Facsymphony a5 flow cytometer, by BD (1 mentions)
Cd19 percp cy5.5 hib19, by BioLegend (1 mentions)
Cd56 pe cf594 ncam16, by BD (1 mentions)
Ccr7 pe cy7 g043h7, by BioLegend (1 mentions)
Hla dr bv711, by BioLegend (1 mentions)
Cd4 bv786 sk3, by BioLegend (1 mentions)
Cd8 buv496 rpa t8, by BD (1 mentions)
Hit8a, by BioLegend (1 mentions)
Gzmb gb11, by BioLegend (1 mentions)
Hit3a, by BioLegend (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
2 protocols using tcr gd fitc b1
1
Multi-panel Flow Cytometry of Immune Cells
2
Comprehensive Immunophenotyping of T-cells
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Extracellular staining was performed to evaluate purity, double positive for CD3-phicoeritrin (Hit3a, BioLegend), CD8-FitC (Hit8a, BioLegend), and TCRg/d-FitC (B1, BioLegend). The naive state of the cells was evaluated by low CD45RO (UCHL1, Tonbo) and by high CD45RA (T6D11, Miltenyi Biotec). For cytokine staining, we used IL-2-APC (MQ1-17H12BD, Pharmingen) and Miltenyi Biotec) . To quantify TLR5 (85B152.5, Abcam), extracellular staining was performed. For cytotoxicity markers, GZMB (GB11, BioLegend) and CD107a (H4A3, Miltenyi Biotec) intracellular staining was performed. For assays of cell division, PBMCs were specifically evaluated by double staining with CFSE (21888, Sigma) 50 nM and CD8-Alexa Fluor 405 (MHCD0826, Life Technologies) or only CFSE in purified CD8 + T cells. For assays of oxidative burst, non-fixed CD8 + T cells were stained with 50 nM dihydroethidium (D7008, Sigma) for 20 min at 37 C, then analyzed immediately on the cytometer.
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