Anti yap taz sc 101199

Cells cultured on coverslips were fixed with 4% paraformaldehyde for 30 min. The cultures were then washed 3 times with PBS, permeabilized with 0.5% Triton X-100 in PBS for 10 min, blocked with 10% BSA and 0.1% Triton X-100 in PBS for 1 h, incubated with primary antibodies (overnight, 4°C), and then washed 3 times with PBS followed by incubation with secondary antibodies (1.5 h, room temperature). Phalloidin was added after the secondary antibodies for 15 min, whereas ProLong-DAPI was added after secondary antibodies or Phalloidin for 15 min. Images were acquired using a confocal microscope (FV1200, Olympus). More than 5 fields of view from at least three independent experiments were randomly chosen.
Primary antibodies were anti-YAP/TAZ (sc-101199) was from Santa Cruz. Anti-phospho-Myosin Light Chain 2 (p-MLC2) (CST 3675) was from Cell Signaling Technology. Secondary antibodies were from Beyotime. ProLong-DAPI was from Invitrogen. Phalloidin (PHDH1) was from Cytoskeleton.

The following antibodies were used: anti-YAP/TAZ (sc-101199, Santa Cruz Biotechnology), anti-tensin-1 (SAB4200283, Sigma-Aldrich), anti-fibronectin (F3648, Sigma-Aldrich), anti-vinculin (clone hVIN-1, V9131), anti-talin (clone 8d4, T3287, Sigma-Aldrich), anti-paxillin (612405, BD Biosciences), anti-phosphotyrosine (Y118)-paxillin (2541, Cell Signaling Technology), anti-phosphotyrosine (Y397)-FAK (clone D20B1, 8556, Cell Signaling Technology) and anti-tensin-3 (ABT29, Millipore). Anti-human active α5-integrin (SNAKA51) was a gift from Martin Humphries, University of Manchester, UK (Clark et al., 2005 (link)). Phalloidin Atto 647N (65906) was obtained from Sigma-Aldrich. Alexa Fluor^®-conjugated secondary antibodies (Alexa Fluor 488, 555, 568, 647-conjugated anti-mouse-, rabbit- and rat-IgG antibodies, Thermo Fisher Scientific) were used in immunofluorescence. Bovine plasma fibronectin (FN; 341631) was purchased from Merck, Millipore and collagen type I (from calf skin) from Sigma-Aldrich (C8919).