Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

EGF Receptor (15F8) Rabbit mAb (IHC Preferred) #4405

Applications Reactivity Sensitivity MW (kDa) Isotype
W IHC-P IF-P H Endogenous 175 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-P=Immunofluorescence (Paraffin)
Reactivity Key:  H=Human
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

EGF Receptor (15F8) Rabbit mAb (IHC Preferred) detects endogenous levels of EGF receptor proteins. The antibody does not recognize other proteins of the ErbB family.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to residues near the carboxy terminus of human EGF receptor.

Western Blotting

Western Blotting

Western blot analysis of various cell lysates using EGF Receptor (15F8) Rabbit mAb (IHC Preferred). This antibody does not cross-react with ErbB2, ErbB3 or ErbB4 expressed in MDA-453, MCF7 and 293/ErbB4 cells respectively.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using EGF Receptor (15F8) Rabbit mAb (IHC Preferred).

IF-P

IF-P

Immunofluorescent analysis of paraffin-embedded human breast carcinoma using EGF Receptor (15F8) Rabbit mAb (IHC Preferred) (green) and DAPI (blue).


Background

The epidermal growth factor (EGF) receptor is a 170 kDa transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for c-Cbl, an adaptor protein that leads to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated residues (Tyr1148 and Tyr1173) provides a docking site for the SHC scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutations to either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

  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.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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.

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