As previously described [4 (link), 14 (link), 16 (link)], total cell lysates were collected from MEFs cultured on polyacrylamide hydrogels by incubating the hydrogels face down for 1 to 2 min at room temperature on 5× SDS sample buffer containing β-mercaptoethanol. Equal amounts of extracted protein were fractionated on reducing 10% SDS-polyacrylamide gels, and the fractioned proteins were subsequently transferred electrophoretically onto nitrocellulose blotting membranes. These membranes were probed with antibodies against FAK (cat. no. 610087, BD Transduction), p130Cas (cat. no. 610271, BD Transduction), or GAPDH (cat. no. sc-25778, Santa Cruz Biotechnology). Immunoblot signals were detected using enhanced chemiluminescence.
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Quantification of Focal Adhesion Proteins
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Other organizations : University at Buffalo, State University of New York, University of Rochester, Cardiovascular Institute Hospital
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Variable analysis
independent variables
- Polyacrylamide hydrogels
- Protein expression levels of FAK
- Protein expression levels of p130Cas
- Equal amounts of extracted protein
- Positive control: GAPDH antibody used for immunoblotting
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