Flow cytometry was conducted using an LSRFortessa (BD Biosciences). A panel consisting of antibodies conjugated to six different fluorophores was used to classify subsets of memory T cells and for drop-seq. Antibodies used were: CD8α-Pacific blue (BioLegend), CD3-BV650 (BD Biosciences), CD45RA-APC-Cy7 (BioLegend), CCR7–488 (Bio-Legend), IL-7Rα-PE (BioLegend) and CD27-PE-Cy7 (BioLegend). For characterization of CSF cells, this same panel was used, but CD19-PE-Cy5 (BioLegend) and CD14-Qdot-705 (Thermo Fisher Scientific) were included. For sorting CSF T cells for TCR plate-seq, the following antibodies were used: CD8α-PE (BioLegend), CD161-PE-Cy7 (BioLegend), CXCR3-APC (BioLegend), CD4-APC-Alexa700 (Thermo Fisher Scientific), CD39-APC-Cy7 (BioLegend), CD38-FITC (BioLegend), PD-1-BV421 (BD Biosciences), CD45RA-BV605 (BD Biosciences), CD3-BV650 (BD Biosciences), CD27-BV786 (BD Biosciences) and CD127-BUV395 (BD Biosciences). For each experiment, a compensation matrix was developed using singly stained and unstained controls or fluorescent beads, and all analysis was conducted in Cytobank.
Cd127 buv395
Manufactured by BD
CD127-BUV395 is a fluorochrome-conjugated antibody that binds to the CD127 antigen. CD127 is a surface marker expressed on various cell types, including lymphocytes. The BUV395 fluorochrome allows for detection and analysis of CD127-expressing cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Cd8α pacific blue, by BioLegend (1 mentions)
Pd 1 bv421, by BD (1 mentions)
Cd38 fitc, by BioLegend (1 mentions)
Cd27 pe cy7, by BioLegend (1 mentions)
Cd19 pe cy5, by BioLegend (1 mentions)
Cd45ra apc cy7, by BioLegend (1 mentions)
Cd8α pe, by BioLegend (1 mentions)
Cxcr3 apc, by BioLegend (1 mentions)
Cd27 bv786, by BD (1 mentions)
Cd45ra bv605, by BD (1 mentions)
Cd3 bv650, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
Cd4 pe, by Beckman Coulter (1 mentions)
Cd45ra fitc, by BD (1 mentions)
Cxcr3 bv711, by BD (1 mentions)
Ccr6 percp cy5, by BD (1 mentions)
Cd25 bv786, by BD (1 mentions)
Ficoll density centrifugation, by Merck Group (1 mentions)
Ccr7 pe, by BD (1 mentions)
Facscelesta, by BD (1 mentions)
2 protocols using cd127 buv395
1
Multiparameter Flow Cytometry of CSF Immune Cells
2
Multiparametric Characterization of CD4+ T Cells
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Peripheral blood mononuclear cells (PBMCs) were obtained after Ficoll density centrifugation (Sigma-Aldrich, Burlington, MA). CD4+ T cells were purified from blood using negative selection on magnetic columns (CD4 T-cell isolation kit, human; Miltenyi Biotec, Bergisch Gladbach, Germany). Cells were stained using the following antihuman antibodies: CXCR5-BV605, CD45RA-FITC, CCR7-PE, CXCR3-BV711, CCR6- PercpCy5.5, PD-1-BUV737, CD25-BV786, and CD127-BUV395 (BD Biosciences, Franklin Lakes, NJ). For CSF samples and transmigration assays, CCR7-PE was replaced by a CD4-PE (Beckman Coulter, Brea, CA). Cells were analyzed by flow cytometry (FACS Celesta, BD Biosciences). Dead cells were excluded from the analysis using live/dead Amcyan staining (Invitrogen, Waltham, MA).
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