Secondary anti rabbit hrp igg antibodies

Cells were lysed in RIPA lysis buffer (Bryotime, Shanghai, China) supplemented with 1× protease inhibitor cocktail (Thermo-Pierce Biotechnology, Rockford, IL) and sonicated on ice. The protein concentration was determined by the BCA protein concentration assay kit (Bryotime, Shanghai, China) and equal amounts of proteins were separated by gel electrophoresis, and then electroblotted to the polyvinylidene difluoride (PVDF) membrane, blocked with 5% non-fat milk, incubated with the primary antibody and the secondary antibody sequentially. Finally, target proteins were detected using ECL reagent (YESEN, Shanghai, China) and photographed by ChemiDoc^TM XRS+ SYSTEM (Bio-Rad, CA, USA). Primary antibodies for β-Actin (ABclonal, Woburn, MA), DHFR (Abcam, Cambridge, UK), CD38-human, CD38-mouse, Flag (Cell Signaling Technology, Danvers, MA), and secondary anti-rabbit HRP-IgG antibodies (Cell Signaling Technology, Danvers, MA) were used for detecting target proteins.