Overview

  • Product name

    Mouse EGFR ELISA Kit
    See all EGFR kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 1.9%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 8.5%
  • Sample type

    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    60 pg/ml
  • Range

    1.875 ng/ml - 120 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 102 97.1% - 106.7%
    Cell culture media 116.1 114.3% - 119.3%
    Heparin Plasma 96.9 95.2% - 99.7%
    EDTA Plasma 101.1 97.5% - 106%
    Citrate Plasma 100.5 96.2% - 105%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Human, Pig
  • Product overview

    Abcam’s EGFR in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of EGFR protein in mouse cell culture supernatant, serum and plasma samples.

    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

    Sensitivity:

    Samples diluted in Sample Diluent NS – 0.06 ng/mL
    Samples diluted in Sample Diluent 25 BS – 0.98 ng/mL

  • Notes

    EGFR is a receptor tyrosine kinase that binds ligands of the EGF family and activate several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. The ligand binding triggers receptor homo- and/or hetero-dimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. EGFR activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLC gamma-PKC and STATs modules. EGFR may also activate the NF-kappa-B signaling cascade. EGFR also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. EGFR also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin. Endocytosis and inhibition of the activated EGFR by phosphatases like PTPRJ and PTPRK constitute immediate regulatory mechanisms. Upon EGF-binding EGFR phosphorylates EPS15 that regulates EGFR endocytosis and activity. Moreover, inducible feedback inhibitors including LRIG1, SOCS4, SOCS5 and ERRFI1 constitute alternative regulatory mechanisms for the EGFR signaling.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse EGFR Capture Antibody 1 x 600µl
    10X Mouse EGFR Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CP 1 x 6ml
    Mouse EGFR Lyophilized Recombinant Protein 1 x 2 vials
    Plate Seals 1 unit
    Sample Diluent 25BS 1 x 20ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 x 96 units
    Stop Solution 1 x 12ml
    TMB Development Solution 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

    • 9030024J15Rik
    • AI552599
    • Avian erythroblastic leukemia viral (v erb b) oncogene homolog
    • Avian erythroblastic leukemia viral (verbb) oncogene homolog
    • Cell growth inhibiting protein 40
    • Cell proliferation inducing protein 61
    • EGF R
    • EGFR
    • EGFR_HUMAN
    • Epidermal growth factor receptor
    • Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog)
    • Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian)
    • Erbb
    • erbb 1
    • Erbb1
    • Erythroblastic leukemia viral (v erb-b) oncogene homolog
    • HER1
    • mENA
    • Oncogene ERBB
    • OTTMUSP00000005385
    • OTTMUSP00000005386
    • PIG61
    • Proto oncogene c ErbB 1
    • Proto-oncogene c-ErbB-1
    • Receptor tyrosine protein kinase ErbB 1
    • Receptor tyrosine protein kinase ErbB1
    • Receptor tyrosine-protein kinase ErbB-1
    • RP23-295E4.1
    • Urogastrone
    • wa2
    • Wa5
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab201275 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

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Standard curve comparison between mouse EGFR SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of EGFR were measured in duplicates, interpolated from the EGFR standard curve and corrected for sample dilution. Note that 1X Diluted serum and plasma sample were 200X pre-diluted samples, The interpolated, dilution factor-corrected values are plotted (mean +/- SD, n=2).

  • The concentrations of EGFR were measured in duplicates, interpolated from the EGFR standard curve and corrected for sample dilution. The interpolated, dilution factor-corrected values are plotted (mean +/- SD, n=2).

Protocols

References

This product has been referenced in:

  • Kyohara M  et al. Serum Quantitative Proteomic Analysis Reveals Soluble EGFR To Be a Marker of Insulin Resistance in Male Mice and Humans. Endocrinology 158:4152-4164 (2017). Read more (PubMed: 29028997) »
See 1 Publication for this product

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