Antibody against β actin clone ac 15

Antibodies against STIM1 (1:1000), Myosin II (1:1000) and phospho-Ser19 Myosin light chain II (1:1000) were purchased from Cell Signaling Technology. Antibody against β-actin (clone AC-15) (1:5000) was purchased from Sigma-Aldrich. Antibodies against MMP2 (1:1000) and MMP9 (1:1000) were purchased from Millipore. Antibodies against MT1-MMP (1:500) and cortactin (1:100) were purchased from Santa Cruz Biotechnology. Thapsigargin, SKF-96365, and 2-APB were from Cayman Chemical. For immunoblotting, cells were harvested with ice-cold modified radioimmune precipitation assay (RIPA) buffer containing a protease inhibitor mixture (Roche Diagnostics), 100 mM KCl, 80 mM NaF, 10 mM EGTA, 50 mM h-glycerophosphate, 10 mM p-nitrophenyl phosphate, 1 mM vanadate, 0.5% sodium deoxycholate, and 1% NP40. Protein concentrations were determined with the use of a Bio-Rad protein assay. Equal amounts of protein lysates were separated by SDS-PAGE, and then transferred to nitrocellulose blotting membranes (Pall). Immunoblots were blocked, incubated with the primary antibody, washed, and incubated with the corresponding horseradish peroxidase-conjugated secondary antibody (Jackson ImmunoResearch), and visualized by Western blotting luminol reagent (Santa Cruz Biotechnology). Bands in the immunoblots were quantified using Vision WorksLS software (UVP).