Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.88 pg/ml
- Range: 9.4 pg/ml - 600 pg/ml
- Sample type: Cell culture extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Product nameHuman EphA2 ELISA Kit
See all Eph receptor A2 kits
Intra-assay Sample n Mean SD CV% Overall 8 2.6% Inter-assay Sample n Mean SD CV% Overall 3 4.2%
Sample typeCell culture extracts
Assay typeSandwich (quantitative)
Range9.4 pg/ml - 600 pg/ml
Sample specific recovery Sample type Average % Range Cell culture extracts 108 105% - 113%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Human EphA2 ELISA Kit (ab219051) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of EphA2 protein in cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human EphA2 with 0.88 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage instructionsStore 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
FunctionReceptor 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 specificityExpressed 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 diseaseGenetic 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 similaritiesBelongs 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.
modificationsActivated 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.
- Information by UniProt
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.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab219051 has not yet been referenced specifically in any publications.