Anti tmprss2 antibody

HepG2 or 293 T cellular extracts after indicated treatments were collected and prepared according to previously studies for Western blot analysis [19 (link),20 (link)]. Equal amounts of protein in each groups were fractionated on a 10 % SDS-PAGE and transferred to polyvinylidene difluoride membranes. Then, the membranes were blocked by 5% nonfat dried milk for 30 min. The membranes were incubated in primary indicated antibody for 12 h at room temperature. These primary antibodies included: anti-ACE2 antibody (Cell Signaling, ratio: 1:1000), anti-TMPRSS2 antibody (Abcam, ratio: 1:1000) and anti-GAPDH antibody (Santa Cruz, IB: 1:10000). Both the primary antibodies and the secondary antibodies were diluted with 1% nonfat dried milk in 0.1 % TBST (Tris-Buffered Saline Tween-20). Next, the membranes were washed by 0.1 % TBST and incubated in horseradish peroxidase-conjugated secondary anti-mouse or anti-rabbit antibodies (Santa Cruz, ratio: 1:5000) which were diluted with 1% nonfat dried milk in 0.1 % TBST for one hour at room temperature. The protein signals were detected by chemiluminescence through the Super Signal substrate (Pierce, catalog number: 34,087).