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Lysates from untreated EGFP-Paxillin β-transfected Hek293 cells (1, 4), lysates from EGFP-Paxillin β-transfected Hek-293 cells treated with EGF (2, 5), and lysates from EGFP-Paxillin S178A mutant tranfected Hek-293 treated with EGF (3, 6), were resolved on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% milk-TBST buffer for one hour at room temperature, and then were incubated with the Paxillin [pS178] antibody (1-3) or anti-Paxillin pan antibody (4-6) for two hours at room temperature in a 1% milk-TBST buffer. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate, and bands were detected using the Pierce SuperSignal™ method.
![Lysates from untreated EGFP-Paxillin β-transfected Hek293 cells (1, 4), lysates from EGFP-Paxillin β-transfected Hek-293 cells treated with EGF (2, 5), and lysates from EGFP-Paxillin S178A mutant tranfected Hek-293 treated with EGF (3, 6), were resolved on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% milk-TBST buffer for one hour at room temperature, and then were incubated with the Paxillin [pS178] antibody (1-3) or anti-Paxillin pan antibody (4-6) for two hours at room temperature in a 1% milk-TBST buffer. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate, and bands were detected using the Pierce SuperSignal™ method.](http://tools.thermofisher.com/content/sfs/gallery/low/3752.jpg)