MBTP 32: Immunophenotyping of human γδ T cells using flow cytometry

Miltenyi Biotec-tested panel 32 (MBTP32)

This application protocol describes the analysis of differentiation stages of human γδ T cells in peripheral blood mononuclear cells (PBMCs) using flow cytometry. 

Protocol

Figure 1: Gating strategy showing the analysis of γδ T cells from human PBMCs. Samples were initially gated on lymphocytes based on SSC-A/FSC-A gating (A). Single cells were gated using FSC-H/FSC-A and SSC-A/SSC-H gating (B, C). Cells were further gated on live cells using 7-AAD-negative gating (D). T cells were gated as CD3+ (E). From total CD3+ T cells, γδ T cells were characterized based on their lack of CD4 and CD8 expression and high expression of TCRγ/δ (F, G). TCRγ/δ+ T cells (γδ T cells) were further discriminated into TCR Vδ1+, TCR Vδ2+, and TCR Vδ1 TCR Vδ2 subpopulations (H). 

Figure 2: Gating strategy showing the analysis of TCR Vδ1+ T cells. Samples were initially gated as shown in Figure 1. The differentiation phenotype of TCR Vδ1+ T cells was characterized upon expression of CD27, CD28, CD45RA, and CD16 (A-D).

Figure 3: Gating strategy showing the analysis of TCR Vδ2+ T cells. Samples were initially gated as shown in Figure 1. The differentiation phenotype of TCR Vδ2+ T cells was characterized upon expression of CD27, CD28, CD45RA, and CD16 (A-D).

Materials

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