Human apj apc conjugated antibody

The cells were stained with monoclonal antibodies (mAb) and analyzed using a FACS Verse (Becton Dickinson, Franklin Lakes, NJ). The following mAbs were used for the phenotypic analysis: eFluor450‐labeled anti‐CD3 (eBiosciences, San Diego, CA, 48‐0037‐41), PE‐Cy7‐labeled anti‐CD3 (BD PharMingen, San Diego, CA, 563423), FITC‐labeled anti‐TCR Vγ9 (Beckman Coulter, Brea, CA, IM1463), Human APJ APC‐conjugated antibody (R&D systems, Minneapolis, MN, FAB856A), APC anti‐human CD34 (Biolegend, San Diego, CA, 343608) and Anti‐Human CD43 PE (eBioscience, 12‐0439).
On days 4 and 12 in hematopoietic differentiation, the cells were treated with 500 µl of Accutase at 37°C for 15 minutes and washed with 1% BSA in PBS after centrifuging, followed by reactions with antibodies. On day 12, the dissociated cells were resuspended in StemPro34 medium and plated onto plastic at 37°C for 1 hour, and the floating cells were collected to remove any adherent cells. Dead cells were excluded by 7‐AAD staining.

Cells were pretreated with Accutase (Invitrogen) for 3 minutes at 37°C and then washed with cold PBS. After 300×g centrifugation, 2×10⁵ cells were resuspended in 100 ml of PBS and incubated with primary antibodies for 30 minutes at 4°C. Then, the cells were washed by using PBS with 2% FBS and 0.2 mM EDTA and used for flow cytometry analysis or sorting. The antibodies were as follows: Human APJ-APC-conjugated Antibody (R&D), PE Mouse Anti-Human CD31 (BD Biosciences), APC Mouse Anti-Human CD34 (BD Biosciences), APC Mouse Anti-Human CD43 (BD Biosciences), Mouse IgG3 APC-conjugated Antibody (R&D), PE Mouse IgG1, κ Isotype Control (BD Biosciences).