Rabbit anti vp1

Cells were lysed with SAM buffer (50 mM Tris-HCl, pH 6.8, 3% of SDS, 5% of β-mercapto-ethanol) containing protease inhibitor cocktail (SIGMAFAST Protease Inhibitor Tablets; Sigma-Aldrich), resolved by SDS-PAGE (10% polyacrylamide gels under reducing conditions) and electro-transferred to nitrocellulose membranes (Bio-Rad Laboratories). Membranes were blocked with 5% nonfat milk and probed with different primary antibodies for 1 hr at room temperature: (1) mouse anti-SV40 LTag 0.5 mg/mL (BD Biosciences), dilution 1:200; (2) rabbit anti-VP1 (Abcam), dilution 1:5,000; (2) mouse anti-GFP (Sigma-Aldrich), dilution 1:1,000; mouse anti-α-tubulin antibody (Sigma-Aldrich), dilution 1:5,000; mouse anti-GAPDH (Millipore), dilution 1:5,000. Finally, the membrane was hybridized with a goat anti-mouse or a goat anti-rabbit antibody conjugated with horseradish peroxidase (HRP) (Sigma-Aldrich), dilution 1:5,000. The blot was developed by chemiluminescence (Chemidoc MP Imaging System; Bio-Rad Laboratories). To quantify the amounts of protein, we scanned the blots by densitometry using the Image Lab software version 5.2.1 (Bio-Rad Laboratories).