Mouse monoclonal anti zeb2

The protein was extracted by radioimmunoprecipitation assay lysis buffer from indicated cells, and BCA Protein Assay Kit (Thermo Scientific) was used to measure the protein concentration. After being separated on 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis gels, the protein was transferred to nitrocellulose membranes. The membranes were blocked with 5% nonfat dry milk and incubated with primary antibodies (rabbit polyclonal anti-E-cadherin [1:1,000], mouse monoclonal anti-vimentin [1:1,000], rabbit polyclonal anti-fibronectin [1:1,500], and rabbit monoclonal anti-beta Catenin [1:2,000] from Abcam, Cambridge, UK; goat polyclona anti-ZEB1 [1:500] and mouse monoclonal anti-ZEB2 [1:500] from Santa Cruz Biotechnology Inc., Dallas, TX, USA; mouse monoclonal anti-GAPDH [1:5,000] from Sigma-Aldrich Co., St Louis, MO, USA) overnight at 4°C. The membranes were washed with TBST, and then incubated with appropriate horseradish peroxidase-conjugated secondary antibody. Enhanced chemiluminescence reagent was used to detect the signal on the membrane.