Sds kcl precipitation assay kits

Etoposide (VP-16,213) and Doxorubicin were gifts of the Drug Synthesis and Chemistry Branch, Developmental Therapeutic Program of NCI, NIH. Purified topo IIα, supercoiled pHOT1 DNA, kDNA, and SDS/KCl precipitation assay kits were obtained from Topogen (Port Orange, FL). Primary antibodies for the detection of topo IIα was obtained from Abcam, Cambridge, MA. The nitric oxide donor, propylamine propylamine nonoate (PPNO), was obtained from Cayman Chemicals (Ann Arbor, MI). The nitric oxide donor, S-nitrosoglutathione (GSNO) was purchased from Santa Cruz Biotechnology, Inc (Dallas, TX). A stock solution of PPNO or GSNO was prepared in 0.2 N NaOH and was stored at −80°C. Stock solutions of DOX (in H₂O), VP-16 (in DMSO) were prepared and stored at −80°C. Fresh drug solutions prepared from stock solutions were used in all experiments.

Camptothecin was a gift of the Drug Synthesis and Chemistry Branch, Developmental Therapeutic Program of NCI, NIH. Topotecan hydrochloride was obtained from Cayman Chemicals (Ann Arbor, MI). A stock solution of camptothecin was prepared in DMSO, TPT was dissolved in doubly distilled water, and solutions were stored at −80°C. Ascorbic acid and horseradish peroxidase (HRP) were obtained from Sigma-Aldrich Chemical Company (St. Louis, MO). Ascorbic acid (100 mM) was dissolved in chelex-treated phosphate buffer and the pH of the solution was adjusted to 7.2 and stored at −80°C. Human myeloperoxidase was purchased from Lee Biosolutions (St. Louis, MO). Human topo I, antibody to Human topo I, and SDS/KCl precipitation assay kits were obtained from Topogen (Buena Vista, CO). Primary antibody for the analysis of p53 was obtained from Santa Cruz Biochemicals (Dallas, TX). Primary antibody anti-gamma H2AX, for the detection of DNA double-strand breaks, anti-p21, and anti-β-actin were obtained from Abcam (Cambridge, MA). DMPO was purchased from Dojindo (Rockville, MD) and used without any further purification.