Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb #4407

Applications Reactivity MW (kDa) Source Isotype
W IP IHC-P IF-IC H M (R) 175 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb detects endogenous EGF receptors only when phosphorylated at Tyr1173. This antibody may weakly cross-react with insulin receptor.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr1173 of human EGF receptor.

Western Blotting

Western Blotting

Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb specifically binds to tyrosine phosphorylated EGF receptors but not other phosphorylated tyrosine kinases. Western blot analysis of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-EGF Receptor (Tyr1173)(53A5) Rabbit mAb (upper) or Phospho-Tyrosine mAb (P-Tyr-100) #9411 (lower).

Western Blotting

Western Blotting

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 Iressa® (10 uM for 2 hrs), using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb (upper) or EGF Receptor Antibody #2232 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or calf-intestinal phosphatase (CIP) treated (right), using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded MDA-MB-468 cells, untreated (left) or EGF-treated (right), using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using Phospho-EGF Receptor (Tyr1173)(53A5) Rabbit mAb (left) or the same antibody preincubated with Phospho-EGF Receptor (Tyr1173) Blocking Peptide (#1175) (right).


IF-IC

IF-IC

Immunofluorescent staining of A431 cells, untreated (left) or EGF-stimulated (50 ng/ml for 2 min, right), using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of A431 cells either serum-starved (left) or EGF-treated (right) using Phospho-EGF Receptor (Tyr1173) (53A5) Rabbit mAb (green). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

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 autokinase activity (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.

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