ab126441 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphorylated EGFR protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-EGFR (Y1086) and total EGFR (help normalize the results of phospho-EGFR from different cell lysates being compared). An anti-EGFR antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and EGFR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phosphorylated EGFR (Y1086) or anti-total EGFR antibody is used to detect phosphorylated or non-phosphorylated EGFR. After washing away unbound antibody, HRP-conjugated anti-Rabbit IgG or HRP-Streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of EGFR (Y1086) or total EGFR bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Receptor for EGF, but also for other members of the EGF family, as TGF-alpha, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30 and vaccinia virus growth factor. Is involved in the control of cell growth and differentiation. Phosphorylates MUC1 in breast cancer cells and increases the interaction of MUC1 with SRC and CTNNB1/beta-catenin. Isoform 2 may act as an antagonist of EGF action.
Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
Involvement in disease
Defects in EGFR are associated with lung cancer (LNCR) [MIM:211980].
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily. Contains 1 protein kinase domain.
Phosphorylation of Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occur.
Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Co-localizes with TNK2 on the endosomes.
The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of EGFR. Serial dilutions of lysates were analyzed in this ELISA.
Western blot - EGFR (pY1086) + total EGFR ELISA Kit (ab126441)
The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of EGFR. Serial dilutions of lysates were analyzed by Western blot. Immunoblots were incubated with anti-phospho-EGFR (Y1086).