Sds polyacrylamide gel electrophoresis sds page

Cells transduced with individual AAV serotypes were lysed using PRO-PREP^TM Protein Extraction Solution (iNtRON Biotechnology, Seongnam, South Korea). Protein concentrations were determined using the Bio-Rad Protein Assay (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Protein samples (30 μg) were resolved by SDS-polyacrylamide gel electrophoresis (SDS-PAGE; Sigma-Aldrich) on a 10% gel. The resolved proteins were transferred onto membranes and blocked with 5% skim milk (BD, Sparks, MD, USA) for 1 h at room temperature. The membranes were probed for ~12 h using anti-GFP antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Membranes were further probed with antibodies against HSP90 (Santa Cruz Biotechnology), which served as a loading control.

PEG, HPMA, curcumin, acetone, dimethyl formamide (DMF), dimethyl sulfoxide (DMSO), Ficoll-Paque solution, Tris (hydroxymethyl) aminomethane hydrochloride (Tris-HCl), potassium chloride (KCl), 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT), RIPA buffer (containing 50 mM Tris-HCl, 150 mM sodium chloride (NaCl), 1% Triton X-100, 0.5 mM ethylenediaminetetraacetic acid, 0.1% sodium dodecyl sulfate, and protease inhibitor cocktail), SDS-polyacrylamide gel electrophoresis (SD-SPAGE), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were from Sigma-Aldrich (St. Louis, MO, USA). K562, a potent leukemia cell line derived from chronic myelogenous leukemia patients in blast crisis, was purchased from the RIKEN BRC Cell Bank (Ibaraki, Japan). RPMI 1640 medium, fetal bovine serum, penicillin, and streptomycin were from Invitrogen™ Life (Carlsbad, CA, USA). Propidium iodide (PI) was from US Biological (Swampscott, MA, USA). Goat anti-rabbit IgG conjugated with HRP was from the Promega Corporation, (Madison, WI, USA). Luminata^TM Forte Western HRP Substrate was from the Millipore Corporation, (Billerica, MA, USA). Primary rabbit polyclonal anti-WT1 and rabbit polyclonal anti-GAPDH were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Other chemicals and solvents were of the highest grade available.