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Technical Data Sheet


Paxillin Phospho-Regulation
Immunocytochemistry Kit


Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
PM1071 Omori, K. et al. (2015) Bioch Biophys Res Com 458(4): 934. IF: mouse NIH3T3 fibroblasts
PP1341 Wu, D. W. et al. (2014) Oncogene 33(35): 4385. WB: CCM3 cells
PM1071 Cheng, I. et al. (2014) J Biol Chem. 289(39):26989 IF: B16-F1 cells
PM1071 Liu, H. et al. (2013) BMC Cancer. 13:80. WB: human ovarian cancer cells
PM1071 Lee, S. et al. (2013) Biochim Biophys Acta. 1830(8):4017. WB: rat C6 and human U373 gliomas
PP1341 Sen, A. et al. (2012) J Clin Invest. 122(7):2469. WB, ICC: C4C2, PC3, LNCaP
PP1341 Kwak, T.K. et al. (2010) J Biol Chem. 285(46):36021. WB: pancreatic INS-1 cells
PP1341 Sen, A. et al. (2010) J Biol Chem. 285(37):28787. WB: human LNCAP cells
Kit Summary
The paxillin phospho-regulation kit can be used for immunocytochemical co-localization of paxillin phosphorylation at Ser-83 compared to the total expression of paxillin. The kit includes rabbit polyclonal and mouse monoclonal antibodies along with Goat-anti-rabbit conjugated to DyLight® 594 and Goat anti-mouse conjugated to DyLight® 488 for dual labeling experiments.

Immunocytochemical labeling of phosphorylated paxillin in paraformaldehyde fixed and NP-40 permeabilized rat A7r5 cells. The cells were labeled with mouse monoclonal Paxillin (PM1071; left) and rabbit polyclonal Paxillin (Ser-83) (PP1341, right). The antibodies were detected with Goat anti-Mouse DyLight® 488 and Goat anti-Rabbit DyLight® 594, respectively.

Paxillin, a focal adhesion protein, is involved in focal adhesion formation during cell adhesion and migration. Paxillin contains LD motifs, LIM domains, and SH3-/SH2-binding domains that participate in a variety of protein-protein interactions with kinases, GTPase-activating proteins, and cytoskeletal proteins. Phosphorylation of paxillin occurs at both tyrosine and serine sites. Serine phosphorylation of paxillin occurs in response to growth-factor activation and fibronectins. Both ERK and p38MAPK kinases phosphorylate serine 83 in vitro. HGF stimulation of murine epithelial cells leads to ERK-mediated phosphorylation of Ser-83, which is required for HGF-induced cell spreading and migration. In addition, Ser-83 is phosphorylated in response to NGF in PC12 cells, and this phosphorylation may be involved in neurite extension. In human paxillin, Ser-85 rather than Ser-83 may be the site phosphorylated by p38 MAPK and mutation of this site inhibits NGF-induced neurite extension. Thus, serine residues in the N-terminal region of paxillin may be important for growth-factor mediated changes in activity.

Background References
Woodrow, M.A. (2003) Exp. Cell. Res. 287(2):325-338.
Huang, C. et al. (2003) Nature 424:219-223.
Huang, C. et al. (2004) J Cell Biol. 164(4):593-602.
Huang, C. et al. (2004) Cell Cycle 3(1):4-6.
Ishibe, S. et al. (2004) Mol. Cell 16 :257-267.
Buffer and Storage
Mouse monoclonal, rabbit polyclonal, and secondary reagents are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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