Annexin 5 biotin apoptosis detection kit

The murine colon carcinoma cell line, CT26, murine mammary carcinoma cell line, 4T1 (gift of Yang-Xin Fu), and three human colon cancer cell lines HCT116, SW480, and HT29 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) where each lot was STR profiled. All cell lines were authenticated either by ATCC or IDEXX (4T1), tested for mycoplasma, and used at low passage numbers within 6 months. Cells were grown in the following media: RPMI-1640 (CT26); McCoy's 5A (HCT116); minimum essential medium Eagle (SW480); and Dulbecco's modified Eagle's mMedium (HT29). All media were supplemented with 10% FCS, 2 mM glutamine, and penicillin–streptomycin antibiotic, and the cells were incubated at 37 °C in 5% CO₂. RB solution of 10% was provided by Provectus Biopharmaceuticals (Knoxville, TN, USA). Annexin V-Biotin Apoptosis Detection Kit, streptavidin-FITC, and streptavidin-APC were purchased from eBioscience (San Diego, CA, USA). Anti-β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-CRT (Abcam, Cambridge, MA, USA), anti-HSP90, and anti-GAPDH (Cell Signaling Technology, Danvers, MA, USA) antibodies were purchased and used according to the manufacturer's specifications. DAPI, quinacrin dihydrochloride, and chloroquine diphosphate were purchased from Sigma-Aldrich (St. Louis, MO, USA).