Reagents and Antibodies for Cell Signaling

Reagents were purchased as follows: cisplatin from Santa Cruz Biotechnology (Santa Cruz, CA, USA); etoposide and SP600125 from LC Laboratories (Woburn, MA, USA); SB203580, Roscovitine, LY294002, and CHIR99021 from Sigma-Aldrich; Cycloheximide and MG-132 from Millipore (Billerica, MA, USA); Phos-tag acrylamide from Wako Chemicals USA (Richmond, VA). PD184352 was kindly provided by Steven Grant (Virginia Commonwealth University), which was chemically synthesized in house based on the published structure of the drug. cisplatin was dissolved in PBS [30 (link)] and other reagents were dissolved in dimethyl sulfoxide. The following antibodies were used: Cleaved PARP (D64E10), Cleaved Caspase-3 (5A1E), BCL-X_L, BIM, phospho-S64 MCL-1, COX IV (3E11), β-Actin (13E5) and GAPDH from Cell Signaling Technology (Danvers, MA, USA); Noxa (114C307.1) from Thermo Fisher Scientific (Waltham, MA, USA); MCL-1 from Enzo Life Sciences (Farmingdale, NY, USA) and Rockland Immunochemicals (Gilbertville, PA, USA); BAK (06-536) from Millipore; BAX (N-20), alpha-Tubulin (sc-8035), Myc (9E10) and CDK1 (sc-54) from Santa Cruz Biotechnology; CDK2 from BD Biosciences; HA (3F10) from Roche (Indianapolis, IN, USA). [γ-³²P]ATP was purchased from PerkinElmer (Kanagawa, Japan). Histone H1, recombinant CDK2/Cyclin A, and CDK2/Cyclin E were purchased from SignalChem (Richmond, BC, Canada).