Braf antibody

A375, SKBR3 and NIH-3T3 cells were purchased from the American Type Culture Collection between 2015 and 2018 and grown in the recommended medium. NT-3 cells were obtained from Dr. Jorg Schraeder at the University Medical Center Hamburg-Eppendorf, maintained in RPMI+Glutamax supplemented with 10% fetal bovine serum (FBS), fibroblast growth factor (FGF) and epidermal growth factor (EGF). NT-3 cells were direct sequenced to confirm the absence of mutations in HRAS, KRAS and NRAS. BON cells were acquired from Dr. Mark Hellmich at the University of Texas Medical Branch, and grown in DMEM:F12 medium with 10% FBS. SIG-M5 and JVM3 cells were obtained from Dr. Omar Abdel-Wahab at Memorial Sloan Kettering Cancer Center (MSKCC) and were maintained in RPMI medium with 10% FBS. 3T3 cell lines were engineered to stably express BRAF mutations using a doxycycline-inducible system. Antibodies against ERK, pERK, MEK, pMEK, GAPDH and V5 were obtained from Cell Signaling (Beverly, MA); BRAF antibody was obtained from Santa Cruz Biotechnology (Dallas, TX). EGF and FGF were acquired from Peprotech (Rocky Hill, NJ). PLX4032 (vemurafenib), GSK2118436 (dabrafenib), GSK1120212 (trametinib), and lapatinib were purchased from SelleckChem (Houston, TX). Drugs for in vitro studies were dissolved in dimethyl sulfoxide (DMSO) to yield 10mM or 1mM stock solutions, and stored at -20°C.