Product Pathways - Tyrosine Kinase/ Adaptors
Phospho-p130 Cas (Tyr249) Antibody #4014
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W | H M R | Endogenous | 130 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Protocols
- 4014:
- Western Blotting
Specificity / Sensitivity
Phospho-p130 Cas (Tyr249) Antibody detects endogenous levels of p130 Cas only when phosphorylated at tyrosine 249. The antibody may cross-react with other phosphorylated tyrosines in the substrate domain of p130 Cas.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr249 of human p130Cas. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from NIH/3T3 cells, untreated or calf intestinal phosphatase (CIP)-treated, using Phospho-p130Cas (Tyr249) Antibody (upper) or p130Cas antibody (lower).
Background
p130 Cas (Crk-associated substrate) is a docking protein containing multiple protein-protein interaction domains. The amino-terminal SH3 domain may function as a molecular switch regulating CAS tyrosine phosphorylation, as it interacts with focal adhesion kinase (FAK) (1) and the FAK-related kinase PYK2 (2), as well as with the tyrosine phosphatases PTP-1B (3) and PTP-PEST (4). The carboxy-terminal Src binding domain (SBD) contains a proline-rich motif that mediates interaction with the SH3 domains of Src-family kinases (SFKs) and a tyrosine phosphorylation site (Tyr668 and/or Tyr670) that can promote interaction with the SH2 domain of SFKs (5). The p130 Cas central substrate domain, the major region of tyrosine phosphorylation, is characterized by 15 tyrosines present in Tyr-X-X-Pro (YXXP) motifs, including Tyr165, 249 and 410. When phosphorylated, most YXXP motifs are able to serve as docking sites for proteins with SH2 or PTB domains including adaptors, C-Crk, Nck and inositol 5'-phosphatase 2 (SHIP2) (6). The tyrosine phosphorylation of p130 Cas has been implicated as a key signaling step in integrin control of normal cellular behaviors including motility, proliferation and survival. Aberrant CAS tyrosine phosphorylation may contribute to cell transformation by certain oncoproteins.
- Polte, T. R. et al. (1997) J. Biol. Chem. 272, 5501-5509.
- Astier, A. et al. (1997) J. Biol. Chem. 272, 228-232.
- Liu, F. et al. (1996) J. Biol. Chem. 271, 31290-31295.
- Garton, A. J. et al. (1997) Oncogene 15, 877-885.
- Ruest, P. J. et al. (2001) Mol. Cell. Biol. 21, 7641-7652.
- Bouton, A. H. et al. (2001) Oncogene 20, 6448-6458.
Application References
- Fonseca, P. M. et al. (2004) Regulation and localization of CAS substrate domain tyrosine phosphorylation. Cellular Signalling 16 (5), 621-629. This article references the use of Phospho-p130 Cas (Tyr249) Antibody in the following applications: IC-IF Western Blotting
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Companion Products
- 4011 Phospho-p130 Cas (Tyr410) Antibody
- 4015 Phospho-p130 Cas (Tyr165) Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
