Overview

  • Product name
    Human EGFR ELISA Kit (pS1070)
  • Detection method
    Colorimetric
  • Sample type
    Cell Lysate
  • Assay type
    Semi-quantitative
  • Assay time
    5h 00m
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    ab126434 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 Blot analysis.

    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of phospho-EGFR (Ser 1070) in human cell lysates. An anti-EGFR antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated EGFR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phospho-EGFR (Ser 1070) antibodies are used to detect phosphorylated EGFR. After washing away unbound antibody, HRP-conjugated anti-Rabbit IgG (secondary antibody) 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 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

     

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2x Cell lysate buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    Anti-phospho-EGFR (Ser 1070) 1 x 6µl
    EGFR Microplate (12 strips x 8 wells) coated with monoclonal anti-EGFR 1 unit
    HRP-conjugated anti-rabbit IgG 1 x 25µl
    Positive Control: lyophilized powder from A431 cell lysate 1 vial
    Stop Solution 1 x 8ml
    TMB Substrate Reagent 1 x 12ml
  • Research areas
  • Function
    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.
  • Tissue specificity
    Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
  • Involvement in disease
    Defects in EGFR are associated with lung cancer (LNCR) [MIM:211980].
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    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.
  • Cellular localization
    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.
  • Information by UniProt
  • Alternative names
    • Avian erythroblastic leukemia viral (v erb b) oncogene homolog
    • Cell growth inhibiting protein 40
    • Cell proliferation inducing protein 61
    • EGFR
    • EGFR_HUMAN
    • Epidermal growth factor receptor
    • ERBB
    • ERBB1
    • HER1
    • mENA
    • Oncogene ERBB
    • PIG61
    • Proto-oncogene c-ErbB-1
    • Receptor tyrosine-protein kinase ErbB-1
    • V-ERB-B avian erythroblastic leukemia viral oncogene homolog
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab126434 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • A431 cells were treated with recombinant human EGF at 37°C for 20 min. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phosphoELISA.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed by Western Blot.
  • The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of EGF R. Serial dilutions of lysates were analyzed in this ELISA.
  • The A431 cells were treated with 100 ng/ml recombinant human EGF for 20 minutes to induce phosphorylation of EGF R. Serial dilutions of lysates were analyzed by Western blot. Immunoblots were incubated with anti-phospho-EGFR (Ser1070).

Protocols

References

ab126434 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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