Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-EGF Receptor Antibody Sampler Kit #9922

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
EGF Receptor (D38B1) XP® Rabbit mAb #4267 40 µl W IP IHC-P IF-IC F H M Mk 175 Rabbit IgG
Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 40 µl W IHC-P IF-IC F H M R Mk 175 Rabbit IgG
Phospho-EGF Receptor (Tyr992) Antibody #2235 40 µl W IHC-P H (R) 175 Rabbit
Phospho-EGF Receptor (Tyr1045) Antibody #2237 40 µl W IF-IC H R 175 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each phospho-EGF Receptor antibody recognizes only the phosphorylated form of EGF Receptor at the indicated sites. The control EGF Receptor antibody recognizes both the phosphorylated and nonphosphorylated forms of EGF receptor.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or EGF-treated (100 ng/ml), using Phospho-EGF Receptor (Tyr992) Antibody #2235 (upper) or EGF Receptor Antibody #2232 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or EGF-stimulated (100 ng/ml), using Phospho-EGF Receptor (Tyr1045) Antibody #2237 (upper) or EGF Receptor (C74B9) Rabbit mAb #2646 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts of BxPC-3 cells, untreated or EGF-stimulated, using Phospho-EGF Receptor (Tyr1068) (D7A5) Rabbit mAb #3777 (upper) and EGF Receptor Antibody #2232 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from A-431, BxPC3 and HeLa cells using EGF Receptor (D38B1) Rabbit mAb #4267.

Description

The Phospho-EGF Receptor Antibody Sampler Kit provides an economical means of evaluating the EGF Receptor and several phosphorylation sites that are involved in its activation. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.

Source / Purification

Antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr992, Tyr1045 or Tyr1068 of human EGF receptor. EGF Receptor Antibody is produced by immunizing animals with a fusion protein containing the cytoplasmic domain of human EGF receptor. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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.

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