Polychromatic flow cytometry and cell sorting were performed on stained mononuclear cells as previously described15 (link). Antibodies against the following antigens were used for staining at predetermined concentrations: CD4 (clone OKT4), CD8 (SK1), IFNγ (4S.B3), IL-17 (eBio64DEC17), IL-21 (3A3-N2), IL-22 (IL22JOP), TNFα (MAb11), and PD-1 (J105) from eBioscience; CCR5 (3A9), CD3 (SP34-2), CD8 (SK1), CD16 (HI149), CD20 (2H7), CD23 (M-L233), CD45 (D058-1283), CD123 (7G3), and HLA-DR (L243) from BD; c-Kit (104D2), CD1a (HI149), CD11c (3.9), CD14 (M5E2), CD34 (561), CD95 (DX2), and IL-2 (MQ1-17H12) from Biolegend; CD127 (eBioRDR5) and CD218a (H44) from Thermo; NKp44 (2.9) from Miltenyi; and CD28 (CD28.2) from Beckman Coulter. CD4+ and CD8+ TM were defined as CD95+ singlet, clean, live, CD3+ lymphocytes. NKp44+ ILC3s were defined as CD45+ singlet, clean, live, lineage-negative (CD1a, CD3, CD8, CD11c, CD14, CD16, CD20, CD23, CD34, CD123), NKp44+CD127+CD218+ lymphocytes. Positive/negative gating based on clearly grouped populations, historical-determined expression, and the use of internal controls (Supplementary Fig. 8 ). A threshold of 100 collected events in the parent population was utilized for all subset expression analysis.
Cd127 ebiordr5 and cd218a h44
Manufactured by Thermo Fisher Scientific
CD127 (eBioRDR5) and CD218a (H44) are cell surface markers used for flow cytometry analysis. CD127 is the alpha chain of the interleukin-7 receptor, while CD218a is the interleukin-18 receptor alpha chain. These markers are commonly used to identify and characterize different immune cell subsets.
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2 protocols using cd127 ebiordr5 and cd218a h44
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Polychromatic Cytometry and Cell Sorting
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Polychromatic Cytometry and Cell Sorting
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Polychromatic flow cytometry and cell sorting were performed on stained mononuclear cells as previously described15 (link). Antibodies against the following antigens were used for staining at predetermined concentrations: CD4 (clone OKT4), CD8 (SK1), IFNγ (4S.B3), IL-17 (eBio64DEC17), IL-21 (3A3-N2), IL-22 (IL22JOP), TNFα (MAb11), and PD-1 (J105) from eBioscience; CCR5 (3A9), CD3 (SP34-2), CD8 (SK1), CD16 (HI149), CD20 (2H7), CD23 (M-L233), CD45 (D058-1283), CD123 (7G3), and HLA-DR (L243) from BD; c-Kit (104D2), CD1a (HI149), CD11c (3.9), CD14 (M5E2), CD34 (561), CD95 (DX2), and IL-2 (MQ1-17H12) from Biolegend; CD127 (eBioRDR5) and CD218a (H44) from Thermo; NKp44 (2.9) from Miltenyi; and CD28 (CD28.2) from Beckman Coulter. CD4+ and CD8+ TM were defined as CD95+ singlet, clean, live, CD3+ lymphocytes. NKp44+ ILC3s were defined as CD45+ singlet, clean, live, lineage-negative (CD1a, CD3, CD8, CD11c, CD14, CD16, CD20, CD23, CD34, CD123), NKp44+CD127+CD218+ lymphocytes. Positive/negative gating based on clearly grouped populations, historical-determined expression, and the use of internal controls (Supplementary Fig. 8 ). A threshold of 100 collected events in the parent population was utilized for all subset expression analysis.
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