Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-EGF Receptor Antibody Sampler Kit #9922

Kit Includes Quantity Applications Reactivity MW (kDa) Source
EGF Receptor Antibody # 2232 40 microliters W IP IHC-P IF-IC IC F H M R 175 Rabbit
Phospho-EGF Receptor (Tyr1068) Antibody # 2234 40 microliters W IHC-P H 175 Rabbit
Phospho-EGF Receptor (Tyr992) Antibody # 2235 40 microliters W IHC-P H (M) (R) 175 Rabbit
Phospho-EGF Receptor (Tyr1045) Antibody # 2237 40 microliters W IHC-P IF-IC H 175 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  IC=Immunocytochemistry  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat

Specificity / Sensitivity

Each antibody in this kit detects endogenous levels of the specific site's tyrosinephosphorylated EGF receptor proteins. The control EGF receptor antibody recognizes both the phosphorylated and nonphosphorylated forms of EGF receptor.

Western Blotting

Western Blotting

Western blot analysis of cell lysates from various cell lines, using EGF Receptor Antibody #2232. (A431* cells were treated with EGF 100 ng/ml for 2 minutes.)

Western Blotting

Western Blotting

Western blot analysis of MDA-468 untreated and treated EGF cells, using Phospho-EGF Receptor (Tyr1068) Antibody #2234.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or EGF-stimulated (100 ng/ml) for various times, using Phospho-EGF Receptor (Tyr1045) Antibody #2237 (upper) or EGF Receptor Antibody #2232 (lower).


Western Blotting

Western Blotting

Western blot analysis of A431 cells stimulated with EGF (100 ng/ml) for various times, using Phospho-EGF Receptor (Tyr992) Antibody #2235 (A), Phospho-EGF Receptor (Tyr1045) Antibody #2237 (B), Phospho-EGF Receptor (Tyr1068) Antibody #2234 (C) and control EGF Receptor Antibody #2232 (D).

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-peptides (KLH coupled) corresponding to residues surrounding Tyr845, Tyr992, Tyr1045 or Tyr1068 of human EGF receptor. EGF Receptor Antibody is produced by immunizing rabbits with a synthetic peptide (KLH coupled) corresponding to residues surrounding Tyr1068 of human EGF receptor. Antibodies are purified by protein A and peptide affinity chromatography.

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.

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