Technical Data
P3113-90B
Paxillin (PXN)
Description:
Paxillin can be diffusely distributed in the cytoplasm, co-localized with actin at the cell periphery, or aggregated at focal adhesions. Paxillin has several motifs which allow its interaction with other proteins. These motifs include LD motifs, LIM domains, an SH3 domain binding site, and SH2 domain binding sites. Paxillin complexes with proteins of focal adhesions including vinculin, talin, with the kinases FAK, PYK2, Src, and Csk, with ARF GTPase activating proteins, and with the adaptor protein v-Crk.

Paxillin is phosphorylated at several residues, including tyrosine 31, tyrosine 118, tyrosine 181. The phosphorylation of paxillin appears to be important for several biological functions including S-phase transition and neurite extension.

Hybridoma: Produced by the fusion between BALB/c mouse splenocytes and mouse myeloma NS1 cells.

Applications:
Suitable for use in Immunofluorescence, Western Blotting, Immunoprecipitation, Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Immunofluorescence:
Western Blotting: 1.0ug/ml with incubation for 2 hours at RT
Immunoprecipitation: 2.0 ug/mg of protein lysate
Immunohistochemistry: 1.0-2.0ug/ml, incubation for 30 minutes at RT (on acetone-fixed frozen and formalin-fixed/paraffin-embedded tissue sections)

Staining of formalin/paraffin tissues requires boiling tissue sections in 1 mM EDTA, pH 8.0, for 10-20 minutes followed by cooling at RT for 20 minutes.

Optimal dilutions to be determined by the researcher.

Recommended Positive Control: A431 or HeLa cells. Tonsil tissue. Breast or colon carcinomas.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG12B2.75Affinity Purified
SizeStorageShippingSourceHost
100ug-20CBlue IceHumanMouse
Concentration:
~0.2mg/ml
Immunogen:
Purified recombinant full-length human paxillin protein.
Purity:
Purified from ascites by Protein G affinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.4, 0.2% BSA, 15mM sodium azide
Specificity:
Recognizes paxillin, a protein with Mr68kD. Species Reactivity: Human and rat.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
Cary, L.A. and J.L. Guan (1999) Focal adhesion kinase in integrin-mediated signaling. Front. Biosci. 4:D102-D113.
Grgurevich, S. et al. (1999) The Csk homologous kinase, Chk, binds tyrosine phosphorylated paxillin in human blastic T cells. Biochem. Biophys. Res. Commun. 256(3):668-675.
Murakami, H. et al. (1999) Rho-dependent and -independent tyrosine phosphorylation of focal adhesion kinase, paxillin and p130Cas mediated by Ret kinase. Oncogene 18(11):1975-1982.
Nakamura, K., et al. (2000) Tyrosine phosphorylation of paxillin a is involved in temporospacial regulation of paxillin-containing focal adhesion formation and F-actin organization in motile cells. J. Biol. Chem. 275(35):27155-27164.
Pan, X.Q. et al. (1999) Activation of three classes of nonreceptor tyrosine kinases following Fc gamma receptor crosslinking in human monocytes. Clin. Immunol. 90(1):55-64.
Riedy, M.C. et al. (1999) Activin A and TGF-beta stimulate phosphorylation of focal adhesion proteins and cytoskeletal reorganization in rat aortic smooth muscle cells. Exp. Cell Res. 251(1):194-202.
Schaller, M.D. (2001) Paxillin: a focal adhesion-associated adaptor protein. Oncogene 20(44):6459-6472.
Seko, Y. et al. (1999) Hypoxia induces activation and subcellular translocation of focal adhesion kinase (p125(FAK)) in cultured rat cardiac myocytes. Biochem. Biophys. Res. Commun. 262(1):290-296.
Takayama, Y. et al. (1999) Adenovirus-mediated overexpression of C-terminal Src kinase (Csk) in type I astrocytes interferes with cell spreading and attachment to fibronectin. Correlation with tyrosine phosphorylations of paxillin and FAK. J. Biol. Chem. 274(4):2291-2297.
Thomas S.M. et al. (1999) Characterization of a focal adhesion protein, Hic-5, that shares extensive homology with paxillin. J. Cell. Sci. 112 (Pt 2):181-190.
Weinberg, J.M. et al. (2001) Energetic determinants of tyrosine phosphorylation of focal adhesion proteins during hypoxia/reoxygenation of kidney proximal tubules. Am. J. Pathol. 158(6):2153-2164.