To determine the Treg profile and function, 1x106 PBMCs were incubated for 15 min at room temperature with the anti-CD3-PerCP-Cy™5.5 (clone UCHT1), anti-CD4-APC-H7 (clone SK3), anti-CD8-Alexa Fluor® 700 (clone RPA-T8), anti-CD25-PE-CF594 (clone M-A251), anti-CD127-Alexa Fluor® 647 (clone HIL-7R-M21), anti-CD45RA-PE-Cy™7 (clone HI100), anti-CD39-BV650 (clone TU66), and anti-CD73-BV421 (clone AD2) monoclonal antibodies (all from BD Biosciences). Cells were then washed and fixed/permeabilized (eBioscience, catalog no. 00-5521-00) using FOXP3 staining buffer (eBioscience, catalog no. 00-5523-00), and finally stained with the combination of conjugated antibodies anti-Ki67-FITC (BD Biosciences, catalog no. 556026) and anti-FOXP3-PE (BD Biosciences, clone 259D/C7) for nuclear epitope staining. Next, cells were washed with PBS, fixed with 1% formaldehyde in PBS (Sigma-Aldrich, catalog no. 1004960700) for 15 min at room temperature and analyzed by flow cytometry.
Anti cd45ra pe cy 7 clone hi100
Manufactured by BD
Anti-CD45RA-PE-Cy™7 (clone HI100) is a fluorescently-labeled monoclonal antibody that binds to the CD45RA antigen expressed on human cells. It is designed for use in flow cytometry applications.
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Lab products found in correlation
Anti cd127 alexafluor647 clone hil 7r m21, by BD (1 mentions)
Foxp3 staining buffer, by Thermo Fisher Scientific (1 mentions)
Anti ki67 fitc, by BD (1 mentions)
Anti cd25 pe cf594 clone m a251, by BD (1 mentions)
Anti foxp3 pe, by BD (1 mentions)
Anti tnf α pe cy7, by Thermo Fisher Scientific (1 mentions)
Anti cd4 qdot605, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti cd45ra pe cy 7 clone hi100
1
Comprehensive Treg Immunophenotyping
2
Multi-parameter Immunophenotyping of T Cells
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The following human monoclonal fluorescently-conjugated antibodies were used in this study: anti-CD3 Pacific Blue (clone UCHT1), anti-CD3 APC-H7 (clone SK7), anti-CD4 APC (clone RPA-T4), anti-CD8 PerCP-Cy5.5 (clone SK-1), anti-IFN-γ Alexa Fluor 700 (clone B27), anti-IL-2 FITC (clone 5344.111), anti-IL-2 APC (clone MQ1-17H12), anti-CCR7 FITC (clone 150503), anti-CD45RA PE-Cy7 (clone HI100), all from BD Biosciences, and anti-CD4 QDot605 (clone S3.5; Life Technologies), anti-TNF-α PE-Cy7 (clone Mab11; eBiosciences), and anti-PD-1 PE (clone EH12.2H7; BioLegend). For PD-1/PD-L1 blockade experiments, LEAF™ purified anti-human PD-L1 Ab (clone 29E.2A3) and LEAF™ purified IgG2b isotype-matched control Ab (clone MPC-11) were obtained from BioLegend.
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