Nb200 103

The sample of this study was prepared from celecoxib (Sigma) with 99.995% purity while cisplatin as positive control was prepared from Cisplatin Kalbe. This subject of this study was HeLa cells, a cell culture collection of Parasitology Department, Faculty of Medicine, Universitas Gadjah Mada. The cells were maintained in Dubelcco Modified Eagle Media (Gibco) containing Fetal Bovine Serum (FBS) 10% (v/v) (Gibco) and penicilinstreptomycin 1% (v/v) (Gibco). Cytotoxic assay was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) (Sigma) method while the apoptosis assay was examined by ethidium bromide and acridine orange (Sigma) staining. Propidium iodide reagent contained 0.1% triton X (E.Merck, Germany), 0.2% RNAse and 5% Propidium iodide (Sigma) in Phosphate Buffer Saline (PBS). Immunocytochemistry used p53 primary antibody purchased form Novus Bio NB 200-103, while the universal secondary antibody was derived from Starr Trek Universal HRP Detection System No.901-STUHRP700-090314. All culture plates used in this study were Iwaki® and all tips and micro tubes were supplied by Axigen®.