Anti mouse sirpα clone p84

Cell culture B16-F10 (CRL-6475) cells were obtained from American Type Culture Collection and cultured at 37 °C, 5% CO2 in either RPMI-1640 (Gibco 11835-030) or Dulbecco's Modified Eagle Medium, (DMEM, Corning 10-013-CV) supplemented with 10% (v/v) fetal bovine serum (FBS, Sigma F2442), 100 U/mL penicillin and 100 µg/mL streptomycin (1% P/S, Gibco 15140122). B16 KO cell lines were generated as described previously 1 using single guide RNA (sgRNA) constructs targeting CD47 (5'-TCCCCGTAGAGATTACAATG) and Tyrp1 (5'-CTTGTGGCAATGACAAATTG) and cultured under the same conditions as the parental wild-type B16-F10 cell line. YUMM2.1 cells were a gift from Dr. Chi Van Dang at the Wistar Institute and were cultured under the same conditions in DMEM.
Antibodies used for in vivo treatment and blocking, in vitro phagocytosis, flow cytometry, and Western blotting are as follows: anti-mouse/human Tyrp1 clone TA99 (BioXCell BE0151), isotype IgG2a control clone C1.18.4 (BioXCell, BE0085), anti-mouse CD47 clone MIAP301 (BioXCell BE0270), isotype control rat IgG2a (BioXCell, BE0089), anti-mouse SIRPα clone P84 (BioLegend 144004), isotype control rat IgG1 (BioLegend, 400414). Low-endotoxin and preservative-free antibody preparations were