Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-EGF Receptor (Tyr1148) Antibody #4404

Applications Reactivity Sensitivity MW (kDa) Source
W IHC-P H Mk (R) Endogenous 175 Rabbit

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-EGF Receptor (Tyr1148) Antibody detects endogenous levels of EGF receptor only when phosphorylated at tyrosine 1148. This antibody does not cross-react with other tyrosine-phosphorylated ErbB family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1148 of human EGF receptor. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or EGF-stimulated, using Phospho-EGF Receptor (Tyr1148) Antibody (upper) or EGF Receptor Antibody #2232 (lower).

Western Blotting

Western Blotting

Western blot analysis of various cell lysates containing activated EGFR, ErbB2, ErbB3 and ErbB4 proteins, using Phospho-EGF Receptor (Tyr1148) Antibody (upper) or Phospho-Tyrosine mAb (P-Tyr-100) #9411 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of phosphorylated EGF Receptor in paraffin-embedded human breast carcinoma, showing membrane localization using Phospho-EGF Receptor (Tyr1148) Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded MDA-MB-468 cells, untreated (left), EGF-treated (middle) or EGF and Tarceva®- treated (right), using Phospho-EGF Receptor (Tyr1148) Antibody.

Background

The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, 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 the adaptor protein c-Cbl, leading 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 EGFR residues (Tyr1148 and Tyr1173) provide 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; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

  1. Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
  2. Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
  3. Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-4.
  4. Hubbard, S.R. et al. (1994) Nature 372, 746-54.
  5. Biscardi, J.S. et al. (1999) J Biol Chem 274, 8335-43.
  6. Emlet, D.R. et al. (1997) J Biol Chem 272, 4079-86.
  7. Levkowitz, G. et al. (1999) Mol Cell 4, 1029-40.
  8. Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-66.
  9. Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
  10. Feinmesser, R.L. et al. (1999) J Biol Chem 274, 16168-73.

Application References

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