Overview

  • Product name

    Human EphA2 ELISA Kit
    See all Eph receptor A2 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 2.6%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 4.2%
  • Sample type

    Cell culture extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.88 pg/ml
  • Range

    9.4 pg/ml - 600 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 108 105% - 113%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    EphA2 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of EphA2 protein in cell extracts.


    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.

  • Notes

    Ephrin type-A Receptor 2, EPHA2, is a receptor tyrosine kinase encoded by the gene EPHA2. EphA2 is a 976-amino acid containing transmembrane glycoprotein, and is a member of the ephrin receptor subfamily of the protein-tyrosine kinase family. The extracellular portion of EphA2 contains the ephrin-A ligand binding site, and two fibronectin type-III repeats. EphA2 regulates cell adhesion and differentiation and is thought to be participate in mediating developmental events, especially in the nervous system. EphA2 has been shown to be highly expressed in tumor cell lines.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Reagents

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    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 5BI 1 x 6ml
    10X Human EphA2 Capture Antibody 1 x 600µl
    10X Human EphA2 Detector Antibody 1 x 600µl
    Human EphA2 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Receptor for members of the ephrin-A family. Binds to ephrin-A1, -A3, -A4 and -A5. Plays an important role in angiogenesis and tumor neovascularization. The recruitement of VAV2, VAV3 and PI3-kinase p85 subunit by phosphorylated EPHA2 is critical for EFNA1-induced RAC1 GTPase activation and vascular endothelial cell migration and assembly (By similarity). Induces apoptosis in a p53/TP53-independent, caspase-8-dependent manner.
  • Tissue specificity

    Expressed in brain and glioma tissue and glioma cell lines (at protein level). Expressed most highly in tissues that contain a high proportion of epithelial cells, e.g., skin, intestine, lung, and ovary.
  • Involvement in disease

    Genetic variations in EPHA2 are the cause of susceptibility to cataract cortical age-related type 2 (ARCC2) [MIM:613020]. A developmental punctate opacity common in the cortex and present in most lenses. The cataract is white or cerulean, increases in number with age, but rarely affects vision.
    Defects in EPHA2 are the cause of cataract posterior polar type 1 (CTPP1) [MIM:116600]. A subcapsular opacity, usually disk-shaped, located at the back of the lens. It can have a marked effect on visual acuity.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. Ephrin receptor subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 1 protein kinase domain.
    Contains 1 SAM (sterile alpha motif) domain.
  • Post-translational
    modifications

    Activated by EFNA1 via tyrosine phosphorylation. Phosphorylated residues Tyr-588 and Tyr-594 are required for binding VAV2 and VAV3 while phosphorylated residues Tyr-735 and Tyr-930 are required for binding PI3-kinase p85 subunit. These phosphorylated residues are critical for recruitment of VAV2 and VAV3 and PI3-kinase p85 subunit which transduce downstream signaling to activate RAC1 GTPase and endothelial cell migration. They also play a critical role in transducing EPHA2 signaling in vascular endothelial cells during tumor angiogenesis.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Alternative names

    • ARCC2
    • AW545284
    • CTPA
    • CTPP1
    • CTRCT6
    • EC 2.7.10.1
    • Eck
    • Eph receptor A2
    • EPHA2
    • EPHA2_HUMAN
    • Ephrin receptor
    • Ephrin receptor EphA2
    • Ephrin type A receptor 2
    • Ephrin type-A receptor 2
    • Epithelial cell kinase
    • Epithelial cell receptor protein tyrosine kinase
    • Myk 2
    • Myk2
    • Sek 2
    • Sek2
    • Soluble EPHA2 variant 1
    • Tyrosine protein kinase receptor ECK
    • Tyrosine-protein kinase receptor ECK
    • Tyrosine-protein kinase receptor MPK-5
    • Tyrosine-protein kinase receptor SEK-2
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab219051 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.

     

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

  • The concentrations of EphA2 were measured in duplicate, interpolated from the EphA2 standard curve, and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean EphA2 concentration was determined to be 940 pg/mL in HepG2 cell extract, 1,076 pg/mL in HEK293T cell extract, and 17.4 ng/mL in PC-3 cell extract.

Protocols

References

ab219051 has not yet been referenced specifically in any publications.

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