Overview

  • Product name
    Human soluble EGFR ELISA Kit
    See all EGFR kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Human serum 5 2%
    Inter-assay
    Sample n Mean SD CV%
    Human serum 3 3%
  • Sample type
    Cell culture supernatant, Serum, Cell culture extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    1 pg/ml
  • Range
    78.125 pg/ml - 5000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 95.2 89.1% - 98.9%
    Cell culture media 111 110.3% - 111.8%
    Heparin Plasma 85.8 82.6% - 88.4%
    EDTA Plasma 80.5 65.5% - 88.4%
    Citrate Plasma 83.1 73.8% - 89.3%
    Goat Serum 80 78.3% - 80.9%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s soluble EGFR in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of soluble EGFR protein in human serum, plasma, cell culture supernatant and cell and tissue extract 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 – 1 pg/mL
    Samples diluted in Sample Diluent 1X Cell Extraction Buffer – 10.4 pg/mL

  • Notes

    EGFR is a receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. The 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 heterodimerization 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, PLCgamma-PKC and STATs modules. It 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. It 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 EPS15 is phosphorylated and it regulates EGFR endocytosis and activity. Moreover, inducible feedback inhibitors including LRIG1, SOCS4, SOCS5 and ERRFI1 constitute alternative regulatory mechanisms for the EGFR signaling. EGFR also forms a heterodimer with ERBB2. EGFR is single-pass type I membrane protein of plasma membrane and many intracellular membranes. In response to EGF, EGFR is translocated from the cell membrane to the nucleus via Golgi and ER. EGFR undergo endocytosis upon activation by ligand. Isoform 2 is secreted. EGFR is ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.

  • 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 soluble EGFR Capture Antibody 1 x 600µl
    10X soluble EGFR Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    soluble EGFR Human Lyophilized Recombinant Protein 2 vials
    Stop Solution 1 x 12ml
    TMB Substrate 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 ab193764 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

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

  • 30X diluted serum samples from 10 apparently healthy male donors (1M – 10M) were measured in triplicates using this kit. Interpolated data values are graphed in ng of soluble EGFR per mL of serum (mean +/- SD, n=3). The mean of soluble EGFR concentration of these serum samples was determined to be 68.8 ng/mL with a range of 63.1 – 76.7 ng/mL.

  • Background-subtracted data values (mean +/- SD, n = 2) are graphed.

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

  • 4 molar excess, 2 molar excess and equimolar concentrations of Human recombinant EGF (relatively to molarity corresponding to used soluble EGFR), as well as no EGF were tested in the presence of 2,500 pg/mL Human recombinant soluble EGFR. Same concentrations of EGF were tested also in the absence of soluble EGFR. Interpolated sample values (mean +/- SD, n = 2) are graphed. Note that in the tested range the EGF did not interfere with the soluble EGFR measurement. Also note that the EGF alone did not showed any significant cross-reactivity.

Protocols

References

ab193764 has not yet been referenced specifically in any publications.

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