Fak antibody

Western blot analysis was carried out as previously described (25 (link)). Briefly, tissues and cell lysates were collected, and the protein concentrations were determined using a BCA™ Protein Assay kit (Pierce, Chemical Co., Rockford, IL, USA). Proteins were separated by 10% SDS-PAGE and electro-transferred onto PVDF membranes (Millipore). The membranes were first incubated with the indicated primary antibodies overnight (4°C), followed by HRP-conjugated secondary antibodies for 2 h (room temperature). Blots were visualized using an enhanced chemiluminescence detection kit (Tiangen Biotech Co., Ltd. Beijing, China). The following antibodies were used: mouse monoclonal Kindlin-1 antibody (cat. no. SAB4200465; 1:1,000, Sigma), rabbit monoclonal integrin β1 antibody (cat. no. NB110-57123; 1:1,000, Novus Biologicals, Littleton, CO, USA), mouse monoclonal activated integrin β1 antibody (cat. no. MAB2079Z; 1:1,000, Millipore), rabbit polyclonal FAK antibody (cat. no. 3285; 1:1,000, Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit monoclonal Phospho-FAK (Tyr397) antibody (cat. no. 8556; 1:1,000, Cell Signaling Technology, Inc.), rabbit monoclonal GAPDH antibody (cat. no. 3683, 1:1,000; Cell Signaling Technology, Inc.). All experiments were repeated at least 4 times.