Mouse anti hcv core c7 50

Cells that were cultured in six-well plates in the presence or absence of BBR treatment were lysed with RIPA buffer (Sigma) that was supplemented with cOmplete^TM Tablets Mini Protease Inhibitor Cocktail (ROCHE; Basel, Switzerland) for 30 min on ice. The cells were then clarified at 12000 RPM for 30 min, followed by protein quantitation using the Bio-Rad Protein Assay Kit II (Bio-Rad Laboratories; Hercules, CA, USA). Afterward, the whole cell lysates were separated using SDS-PAGE, and then transferred to a PVDF membrane for probing with the following primary antibodies: rabbit anti-LC3 antibody (Thermo Fisher Scientific) at 1:1000; mouse anti-HCV NS5A (Millipore; MAB8694) at 1:250; mouse anti-HCV core (C7-50) (Thermo Fisher Scientific) at 1:400; and, mouse anti-β-actin (C4) (Santa Cruz Biotechnology; Santa Cruz, CA, USA) at 1:1000. The secondary antibodies that included goat anti-rabbit IgG H&L HRP (Abcam) and anti-mouse IgG HRP (Thermo Fisher Scientific) were used at a 1:3000 dilution. The membranes were finally overlaid with ECL (Bio-Rad) before acquiring the images while using the ChemiDoc-ItTS2 imager (UVP; Upland, CA, USA).