antibody

Epidermal Growth Factor Receptor, phosphorylated (Tyr1173) (erbB, EGFR)

E3375-55

100 ul

monoclonal

domestic rabbit

nonconjugated

phosphorylated

4i5(53A5)

Epidermal Growth Factor Receptor, phosphorylated (Tyr1173) (erbB, EGFR)

rabbit

Mab

Phosphorylated

Antibodies

100 ul

4i5(53A5)

IgG

Epidermal growth factor (EGF) receptor is a 170kD tyrosine kinase. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling and lysosomal degradation (1,2). Phosphorylation of Tyr845 in the kinase domain may stabilize the activation loop, maintain the enzyme in an active state and provide a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of Tyr845 (5). Phospho-tyrosine 992 is a direct binding site for the phospholipase C-g (PLC-g) SH2 domain, resulting in activation of PLC-g-mediated downstreamsignaling (6). Phosphorylation of Tyr1045 creates a major docking site for c-Cbl (7). Binding of c-Cbl to the activated EGF receptor leads to receptor ubiquitination and degradation (8). Phospho-tyrosine 1068 of activated EGF receptor is a direct binding site for Grb2 (9). Phospho-tyrosine 1148 and 1173 provides a docking site for SHC (2). Both sites are involved in activation of MAP kinase signaling. Phosphorylation of EGFR on serine and threonine residues attenuates EGFR kinase activity. Ser1046/1047 in the carboxy-terminal region of EGFR are sites phosphorylated by CaM kinase II. Mutations of Ser1046/1047 upregulate tyrosine autokinase activity of EGFR (10). Applications: Suitable for use in Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested. Recommended Dilution: Western Blot: 1:1000 Immunohistochemistry (Paraffin): 1:250, IHC Protocol: EDTA/TBST Immunoprecipitation: 1:50 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. 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.

Hu Mo Rt

Synthetic phosphopeptide corresponding to residues surrounding Tyr1173 of human EGF receptor.

Recognizes endogenous human EGF receptor only when phosphorylated at tyrosine 1173. May weakly crossreact with insulin receptor. Species Crossreactivity: mouse and rat.

Purified

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

Not determined

Product Type: Mab Isotype: IgG Clone No: 4i5(53A5) Host: rabbit Source: human Concentration: Not determined Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. Purity: Purified Immunogen: Synthetic phosphopeptide corresponding to residues surrounding Tyr1173 of human EGF receptor. Specificity: Recognizes endogenous human EGF receptor only when phosphorylated at tyrosine 1173. May weakly crossreact with insulin receptor. Species Crossreactivity: mouse and rat. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

IHC IP WB

-20°C

1. Hackel, P. O. et al. (1999) Curr. Opin. Cell Biol. 11:184–189. 2. Zwick, E. et al. (1999) Trends Pharmacol. Sci. 20:408–412. 3. Cooper, J.A. and Howell, B. (1993) Cell 73:1051–1054. 4. Hubbard, S. R. et al. (1994) Nature 372:746–754. 5. Biscardi, J. S. et al. (1999) J. Biol. Chem. 274:8335–8343. 6. Emlet, D. R. et al. (1997) J. Biol. Chem. 272:4079–4086. 7. Levkowitz, G. et al. (1999) Mol. Cell 4:1029–1040. 8. Ettenberg, S. A. et al. (1999) Oncogene 18:1855–1866. 9. Rojas, M. et al. (1996) J. Biol. Chem. 271:27456–27461. 10. Feinmesser, R. L. et al., (1999) J. Biol. Chem. 274:16168–16173. 11. Spieker-Polet H, et al., (1995) Proc Natl Acad Sci. Sep 26;92(20):9348-52. 12. Liguori MJ, et al. (2001) Hybridoma. Jun; 20(3):189-98. 13. G.Cano, et al., (2003) Diagn Cytopathol. Oct; 29(4):207 -11. 14. L.K. Diaz and N.Sneige, (2005) Adv Anat Pathol. 12(1):10-19. 15. Z. Huang, et al., (2005) Appl Immunohistochem Mol Morphol. 13(1):91-95. 16. S. Rossi, et al., (2004) Abstract presented at USCAP 2004. Modern Pathology 17(suppl 1):361A. 17. M. Blechner, et al., (2004) Abstract presented at USCAP 2004. Modern Pathology 17(suppl 1):241A. 18. W. Cheuk, et al., (2004) Am J Surg Path. 28(6):801-807. 19. G.B. Budd, et al., (2004) Abstract presented at ASCO Annual meeting, June, New Orleans. 20. S. M. Tarr, et al., (2004) Abstract presented at Association for Molecular Pathology meeting, Los Angeles. 21. A.M. Gown, et al., (2005) Abstract presented at USCAP 2005. Modern Pathology 18(suppl.1):35A. 22. D.O. Treaba, et al., (2005) Abstract presented at USCAP 2005. Modern Pathology 18(suppl.1):53A. 23. S. Rossi, et al., (2005) Am J Clin Pathol. Aug;124(2):295-302.

https://usbio-images.r.worldssl.net/prodimages/24/E3375-55_1.jpg

Western blot analysis of extracts from BxPc-3 cells treated with EGF (100 ng/ml for 5 min) or EGF plus the EGF receptor tyrosine kinase inhibitor Gefitinib (10 uM for 2 hrs), using E3375-55 (upper) or EGF Receptor Antibody (lower).