Mouse anti nsp1 antibody

Western blot was done to study protein levels in whole-cell protein samples or nuclear fractions. Protein samples were loaded onto a SDS polyacrylamide gel. Polyvinylidene difluoride membrane was used for the Western blot as described earlier (Laskar et al., 2011 (link)). The primary antibodies used were mouse anti-Act1 antibody (Abcam), rabbit anti-Rad51 (Abcam), mouse anti-Hsp82 antibody (Calbiochem), rabbit anti-Aha1 antibody (Invitrogen), mouse Anti-DDDDK tag antibody (Abcam), and rabbit anti-GFP antibody (Abcam) at 1:5000 dilutions. For subcellular fractionation, we used mouse anti-Nsp1 antibody (Abcam) as loading control at 1:5000 dilution. For secondary antibodies, horseradish peroxide–conjugated anti-rabbit antibody (Promega) and anti-mouse antibody (Promega) were used at 1:10,000 dilution. The Western blots were developed using chemiluminescent detection system (Thermo Fisher Scientific). Every experiment was repeated at least three times and band intensities were quantified by using ImageJ software. Mean relative densities were plotted using GraphPad Prism.