Caspase 3 fluorescence assay

CS (degree of deacetylation (DDA) 90.28% and molecular weight (MW) 50–200 kDa) was purchased from MP Biomedicals (Eschwege, Germany), HA, R-Alpha-LA, 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS), GSH, Tween 20, Trypsin/EDTA solution 0.25%, rabbit polyclonal antibodies (Abs): CD44, and MTT were provided from Sigma–Aldrich (St. Louis, MO, USA). Fetal Bovine Serum (FBS), L-glutamine (200 mM), and penicillin-streptomycin (10000 U/mL) were obtained from Biochrom (Berlin, Germany). Dead-end Fluorometric TUNEL System and Cell Titer 96^® Aqueous One Solution Cell Proliferation Assay were prepared from Promega (Madison, USA). MCF-7 and BT-20 cell lines were supplied from Pasteur Institute (Tehran, Iran) and MT from Aburaihan Pharmaceutical (Tehran, Iran). Dulbecco’s Modified Eagle Medium (DMEM) and MEM media, Fluorescein isothiocyanate (FITC)-conjugated (green) secondary Ab, and anti-fade reagent were obtained from GIBCO Invitrogen (Grand Island, NY). Kits of LDH cytotoxicity, Caspase-3 Fluorescence assay, and JC-1 Mitochondrial Membrane Potential assay were supplied from (Cayman Chemical, Michigan, MI, USA). All of the other chemicals and solvents were purchased from Merck (Darmstadt, Germany).