Lactate dehydrogenase ldh release detection kit

EVO (98% purity) was purchased by Sigma-Aldrich. DOX (98% purity) was obtained from Meilun Biology Technology Company (Dalian, China). Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin (PS), phosphate-buffered saline (PBS), propidium iodide (PI) and 0.25% w/v trypsin/1 mM EDTA were purchased from Gibco Life Technologies (Grand Island, USA). The lactate dehydrogenase (LDH) release detection kit was obtained from Roche Diagnostics. Hoechst 33342 and 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) were obtained by Molecular Probes (Grand Island, USA). Primary antibodies against cleaved caspase-7, cleaved caspase-9, cleaved PARP, Ras, phosphorylated MEK, MEK, phosphorylated ERK1/2, ERK1/2, XIAP, cIAP1, survivin, P-gp and GAPDH and secondary antibodies were purchased from Cell Signaling Technology.

EVO, paclitaxel (PTX), crystal violet, and 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO). Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin (100 U/ml)-streptomycin (100 μg/ml), phosphate-buffered saline (PBS) and 0.25% w/v trypsin/1 mM EDTA from Gibco Life Technologies (Grand Island, USA) were used for cell culture. Calcium AM and carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) were purchased from Molecular Probes (Grand Island, USA). The lactate dehydrogenase (LDH) release detection kit was purchased from Roche Diagnostics (Mannheim, Germany). The primary and secondary antibodies against Cyclin B1, p27, p21, Rb, p-Cdc2 (Thr161), Cdc2, p-ERK (Thr202/Tyr204), ERK, p38, Ras, GAPDH, and β-actin were purchased from Cell Signaling Technology (Danvers, MA). MDR-1 was obtained from Santa Cruz (Santa Cruz, USA).

Roswell Park Memorial Institute (RPMI) 1640 medium, Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), streptomycin and penicillin were purchased from Life Technologies (Carlsbad, CA, USA). Enzyme-Linked immunosorbent assay (ELISA) kits for IL-1β, IL-2, IL-4, IL-10, IL-12p70, TNF-α and IFN-γ were from R&D Systems (Minneapolis, MN, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was from USB Corp. (Cleveland, OH, USA) and the lactate dehydrogenase (LDH) release detection kit was from Roche Applied Science (Indianapolis, IN, USA). All kits were used according to the manufacturer’s protocols. CTX monohydrate, lipopolysaccharide (LPS), concanavalin (Con)A and dimethyl sulfoxide (DMSO) were from Sigma-Aldrich (St. Louis, MO, USA). YAC-1 cells, a mouse lymphoma cell line, and RAW264.7 cells, a mouse macrophage cell line, were obtained from ATCC (Manassas, VA, USA; no. ATCC^®TIB-160^TM and ATCC^®TIB-71^TM). Antibodies for β-actin, iNOS, NF-κB-p65 and secondary antibody (HRP-linked anti-rabbit IgG) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). piNOS-Luc and pNF-κB-Luc were purchased from Addgene (Watertown, MA, USA).