Technical Data
Epidermal Growth Factor Receptor (erbB, EGFR), phosphorylated (Y1173)
The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands. The epidermal growth factor receptor is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). Mutations affecting EGFR expression or activity could result in cancer.

EGFR (epidermal growth factor receptor) exists on the cell surface and is activated by binding of its specific ligands, including epidermal growth factor and transforming growth factor (TGF ). Upon activation by its growth factor ligands, EGFR undergoes a transition from an inactive monomeric form to an active homodimer. In addition to forming homodimers after ligand binding, EGFR may pair with another member of the ErbB receptor family, such as ErbB2/Her2/neu, to create an activated heterodimer. There is also evidence to suggest that clusters of activated EGFRs form, although it remains unclear whether this clustering is important for activation itself or occurs subsequent to activation of individual dimers.

EGFR dimerization stimulates its intrinsic intracellular protein-tyrosine kinase activity. As a result, autophosphorylation of five tyrosine (Y) residues in the C-terminal domain of EGFR occurs. These are Y992, Y1045, Y1068, Y1148 and Y1173. This autophosphorylation elicits downstream activation and signaling by several other proteins that associate with the phosphorylated tyrosines through their own phosphotyrosine-binding SH2 domains. These downstream signaling proteins initiate several signal transduction cascades, principally the MAPK, Akt and JNK pathways, leading to DNA synthesis and cell proliferation. Such proteins modulate phenotypes such as cell migration, adhesion, and proliferation. The kinase domain of EGFR can also cross-phosphorylate tyrosine residues of other receptors it is aggregated with, and can itself be activated in that manner.

Suitable for use in Western Blot, Immunoprecipitation, ELISA, and Immunocytochemistry.
Other applications not tested.

Recommended Dilution:
Western Blot: 0.5-2ug/ml
Immunoprecipitation: 1-10ug
ELISA: 0.5ug/ml
Immunocytochemistry:1-10ug/ml. TA431 cells were untreated (Panel A) and treated (Panel B) with EGF then stained with 2ug/ml of anti-phospho-EGFR (Tyr1173).
Optimal dilutions to be determined by the researcher.

Immunoblot Analysis:
Lysates from EGF-stimulated and unstimulated HFF cells, 10ug per lane, were resolved by electrophoresis, and transferred to nitrocellulose. EGF-stimulated were probed with E3375-12 and visualized using a rabbit anti-sheep secondary antibody conjugated to HRP. Unstimulated HFF cells were probed with E3375-15 (0.5ug/ml) and visualized using a goat anti-mouse secondary antibody conjugated to HRP. A chemiluminescence detection system was used.

Storage and Stability:
Lyophilized powder may be stored at 4C for short-term only. Reconstitute to nominal volume (see reconstitution instructions for peptides) and store at -20C. Reconstituted product is stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
MabIgG1,k0.T.38Affinity Purified
100ug4C (-20C Glycerol)Blue IceHumanMouse
Synthetic peptide containing the Y1173 motif (NAE[pY 1173 ]LRV) of the human EGF receptor, KLH-conjugated.
Purified by thiophilic adsorption and size exclusion chromatography.
Supplied as a lyophilized powder in PBS, 0.09% sodium azide, PEG, sucrose. Also available without azide. See E3375-15X.
Recognizes the motif NAE[pY]LRV corresponding to the major autophosphorylation site of human and mouse EGF receptor. Does not recognize the non-phosphorylated EGF receptor or other unrelated phosphotyrosine proteins Tyr1173 corresponds to Tyr1197 in the human (isoform a) and mouse precursor proteins, and to Tyr1196 in the mature rat and porcine precursor proteins. Species Crossreactivity: Human, mouse.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Sorkin, A., et al., (1992) J. Biol. Chem. 267:8672-8678. 2. Bertics, P., et al., (1988) J. Biol. Chem. 263:3610-3617.