Anti-Eph receptor A2 antibody [RM-0051-8F21] (ab73254)

Overview

  • Product nameAnti-Eph receptor A2 antibody [RM-0051-8F21]
    See all Eph receptor A2 primary antibodies
  • Description
    Rat monoclonal [RM-0051-8F21] to Eph receptor A2
  • Specificityab73254 is specific for Eph receptor A2. It does not cross react with recombinant mouse Eph receptors A3, 4, 6, 7, 8, and Eph receptors B1, 2, 3, 4, or 6.
  • Tested applicationsSuitable for: IP, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant mouse EphA2 extracellular domain

Properties

Applications

Our Abpromise guarantee covers the use of ab73254 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
IP Use at an assay dependent concentration.
IHC-P 1/100 - 1/200.
Flow Cyt Use 1µg for 106 cells. ab18407-Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

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

Anti-Eph receptor A2 antibody [RM-0051-8F21] images

  • ab73254 at 1/200 dilution, staining Eph receptor A2 in mouse kidney tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).

  • ab73254 Immunoprecipitate in human A431 whole cell lysate. 500µg of cell lysate was incubated with primary antibody (undiluted in 3M ammonium sulphate, 0.1M borate buffer) and matrix (Trosylactivated Dynabeads) for 18 hours at 37°C.
    For western blotting an HRP-Rabbit anti-rat IgG polyclonal (1/500) was used.
    Lane 1: IP with 1500µg protein cell lysate.
    Lane 2: IP with 500µg protein cell lysate.

    See Abreview

  • Overlay histogram showing A431 cells stained with ab73254 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab73254, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-rat DyLigt 488 (IgG  H+L) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG1 [RTK2071] (ab18412, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References for Anti-Eph receptor A2 antibody [RM-0051-8F21] (ab73254)

ab73254 has not yet been referenced specifically in any publications.

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HUES7 ES cells)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 20 µg
Specification HUES7 ES cells
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Username

Abcam user community

Verified customer

Submitted Feb 04 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEPG2)
Specification HEPG2
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 18 2015

Application Western blot
Sample Human Cell lysate - whole cell (HEPG2)
Gel Running Conditions Reduced Denaturing (12.5)
Loading amount 20 µg
Specification HEPG2
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Username

Abcam user community

Verified customer

Submitted Nov 13 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step 1% serum, 0.1% BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 16°C
Sample Mouse Cell (embryonic stem cell)
Specification embryonic stem cell
Permeabilization Yes - 0.1% triton X
Fixative methanol:acetone
Username

Abcam user community

Verified customer

Submitted Apr 04 2014

Application Western blot
Loading amount 1e+006 cells
Gel Running Conditions Non-reduced Denaturing (10% gel)
Sample Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Specification mouse embryonic stem cell
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Username

Abcam user community

Verified customer

Submitted Mar 11 2014

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization No
Sample Mouse Cell (mouse embryonic stem cell)
Specification mouse embryonic stem cell
Gating Strategy live cells in fsc vs ssc
Preparation Cell harvesting/tissue preparation method: cell dissociation buffer
Sample buffer: 0.2% BSA, 0.1%NaAz in PBS
Username

Abcam user community

Verified customer

Submitted Dec 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (A431 cancer cell line)
Total protein in input 500 µg
Specification A431 cancer cell line
Immuno-precipitation step Other - Trosylactivated Dynabeads
Username

Mrs. Alice Bjerregaard

Verified customer

Submitted Dec 18 2009

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