Bv421 conjugated anti human cd45

Cells were labeled with monoclonal antibodies for 30 min at 4 °C in FACS sorting buffer. The following antibodies were used: BV421‐conjugated anti‐human CD45 (BD Biosciences, 563879), FITC‐conjugated anti‐human CD3 (Biolegend, 300306), APC‐H7‐conjugated anti‐human CD4 (BD Biosciences, 560158), Percp‐cy5.5‐conjugated anti‐human CD8 (Biolegend, 344710). After staining, cells were washed once and resuspended in FACS sorting buffer. Cell sorting was performed using BD FACS Aria III. All samples were gated based on forward and side scatters, followed by exclusion of doublets. Expression levels of CD45, CD3, CD4, and CD8 were gated by their negative controls of unstained cells and positive controls of cells stained by each antibody. CD45⁺CD3⁺ T cells were sorted into 1.5 mL tubes containing 300 µL ice‐cold PBS with 1% BSA. The abundance of the relevant cell populations determined by testing the sorted cells again by FACS, reached more than 99%. Data analysis was performed using FlowJo V10 software (http://www.flowjo.com).

Cells were stained in FACS sorting buffer with specific antibodies for 30 min at 4 °C. The following antibodies were used: BV421-conjugated anti-human CD45 (BD Biosciences, 563879), FITC-conjugated anti-human CD34 (BD Biosciences, 555821), APC-Cy7-conjugated anti-human CD235a (BioLegend, 349116), BV605-conjugated anti-human CD44 (BD Biosciences, 562991) and PerCP-Cy5.5-conjugated 7-AAD (eBioscience, 00-6993-50). After staining, cells were washed once and resuspended in FACS sorting buffer. Cells were sorted on BD FACS Aria II. Pre-gating was first done for live cells based on a 7-AAD staining. Data analysis was performed using FlowJo V10 software (https://www.flowjo.com).