HBB-targeted HSPCs were stained for hematopoietic stem and progenitor cell surface markers at days 4 or 12 post-targeting. Cells were spun down at 300 × g for 5 min and resuspended in 100 μL staining buffer containing an antibody mastermix supplied with the following human specific antibodies: 2 μL CD34-APC-Cy7 (clone 561; BioLegend); 2 μL CD45RA-BV605 (clone HI100; BioLegend); 2.5 μL CD90-BV421 (clone 5E10; BD Biosciences); and 5 μL CD201 (EPCR)-APC (BioLegend). Cells were incubated on ice for 30 min in the dark, washed with PBS, and resuspended in FACS buffer containing 1 μg/mL propidium iodide and analyzed on a FACS Aria II SORP.
Cd45ra bv605 clone hi100
Manufactured by BioLegend
CD45RA-BV605 (clone HI100) is a monoclonal antibody conjugate that targets the CD45RA protein. It is designed for use in flow cytometry applications.
Automatically generated - may contain errors
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Intracellular fixation permeabilization buffer set, by Thermo Fisher Scientific (1 mentions)
Cd3 bv785 clone okt3, by BioLegend (1 mentions)
Ccr7 percp cy5.5 clone g043h7, by BioLegend (1 mentions)
Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
2 protocols using cd45ra bv605 clone hi100
1
Hematopoietic Stem Cell Immunophenotyping
2
Comprehensive SARS-CoV-2 PBMC Profiling
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After stimulation with SARS-CoV-2 OPP, PBMCs were stained with fluorophore conjugated antibodies for flow cytometry analysis. Mastermixes, containing all antibodies for intra or extracellular staining in the optimal concentrations were prepared directly before staining. At first, cells were surface stained with CCR7-PerCP-Cy5.5 (clone G043H7) (BioLegend), CD4-A700 (clone OKT4) (BioLegend), Fixable Viability Dye eFluor780 (eBioscience), CD8-V500 (clone RPA-T8) (BD Biosciences) and CD45RA-BV605 (clone HI100) (BioLegend) for 10 minutes at room temperature in the dark. After thoroughly washing with PBS/BSA, cells were fixed and permeabilized using Intracellular Fixation & Permeabilization Buffer Set according to the manufacturer’s instructions (Thermo Fisher Scientific). Then, the cells were stained with Granzyme B-FITC (clone GB11) (BioLegend), IL2-PE (clone MQ1-17H12) (BioLegend), IL4-PE-Dazzle594 (clone MP4-25D2) (BioLegend), CD137 (4-1BB)-PE-Cy7 (clone 4B4-1) (BioLegend), CD154 (CD40L)-A647; (clone 24-31) (BioLegend), TNFa-eFluor450 (clone MAb11) (eBioscience), IFNg-BV650 (clone 4S.B3) (BioLegend), CD3-BV785 (clone OKT3) (BioLegend) for 30 minutes at room temperature in the dark. All samples were washed thoroughly with PBS and acquired on a CytoFlex flow cytometer (Beckman Coulter).
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