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.\n\nEGFR (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 a (TGFa). 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.\n\nEGFR 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.\n\nApplications:\nSuitable for use in Western Blot, Immunoprecipitation, ELISA, and Immunocytochemistry.\nOther applications not tested.\n\nRecommended Dilution: \nWestern Blot: 0.5-2ug/ml \nImmunoprecipitation: 1-10ug \nELISA: 0.5ug/ml\nImmunocytochemistry: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).\nOptimal dilutions to be determined by the researcher.\n\nImmunoblot Analysis: \nLysates 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.\n\nStorage and Stability: \nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume (see reconstitution instructions for peptides) and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. 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.

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SPECIFICATIONS

Catalog Number

E3375-15

Size

50ug

Applications

ELISA, ICC, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse

Form

Supplied as a lyophilized powder in PBS, 0.09% sodium azide, PEG, sucrose. Also available without azide. See E3375-15X.

P Type

Mab

Purity

Purified by thiophilic adsorption and size exclusion chromatography.

Isotype

IgG1,k

References

1. Sorkin, A., et al., (1992) J. Biol. Chem. 267:8672-8678. 2. Bertics, P., et al., (1988) J. Biol. Chem. 263:3610-3617.

Additional Info

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.