Anti vimentin primary antibody

After exposure, the culture medium was sucked and discarded, washed twice with precooled PBS, and the cells were scraped off with a cell scraper and transferred to a centrifuge tube to make the cell suspension. The total protein was extracted with the Whole Cell Lysis Assay (KGP250, KeyGen Biotech Co., Ltd.), and the protein concentration was detected by the BCA method. After SDS-PAGE electrophoresis, it was wet-transferred to the PVDF membrane. After blocking with TBST containing 5% skimmed milk for 1 h, the primary antibody (anti-β-actin antibody, anti-E- cadherin primary antibody, anti-Vimentin primary antibody, ProteinTech, USA) was incubated overnight (the dilution ratio was 1:1000 and the incubation temperature was 4 °C). The next day, the secondary antibody (ProteinTech, USA) was incubated for 1 h (the dilution ratio was 1:2000 and the incubation temperature was 4 °C). After that, the membrane was washed with TBST solution 3 times. Finally, the chemiluminescence imaging system (iBright CL750, Thermo Fisher Scientific, USA) was used for exposure, and the gray value was measured by Image J v1.8.0 software. The ratio of the gray value of the target protein to the gray value of the internal reference protein was calculated as the relative expression of the target protein.