Anti-Paxillin (phospho Y31)抗体(ab4832)

Extracts prepared from NMµMG cells transfected with either the wild-type EGFP-tagged paxillin or a site directed mutant, in which tyrosine 31 was changed to phenylalanine, were resolved on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with 1% BSA, followed by incubation with 0.5 µg/mL anti-paxillin [pY31] antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected by chemiluminescence using the Tropix WesternStar detection method and Kodak BioMax ultraclear film. The data show detection of paxillin phosphorylation with the wild-type but not the Y31F mutant recombinant proteins. Note that the recombinant EGFP-tagged paxillin proteins run at a higher Mr as opposed to the endogenous protein and that in both cell types that the antibody recognizes the endogenous wild-type paxillin protein, which migrates at the expected Mr of 68 kDa.

Extracts prepared from NMµMG cells transfected with either the wild-type EGFP-tagged paxillin or a site directed mutant, in which tyrosine 31 was changed to phenylalanine, were resolved on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with 1% BSA, followed by incubation with 0.5 µg/mL anti-paxillin [pY31] antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected by chemiluminescence using the Tropix WesternStar detection method and Kodak BioMax ultraclear film. The data show detection of paxillin phosphorylation with the wild-type but not the Y31F mutant recombinant proteins. Note that the recombinant EGFP-tagged paxillin proteins run at a higher Mr as opposed to the endogenous protein and that in both cell types that the antibody recognizes the endogenous wild-type paxillin protein, which migrates at the expected Mr of 68 kDa.

Colocalization of phospho Paxillin and actin in TGF-beta treated NMµMG cells. Green = actin stress fibers, Orange = Paxillin [pY31], Yellow = colocalization.