Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y604) antibody (ab61791)

Overview

  • Product name
    Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y604) antibody
    See all Eph receptor B1 + Eph receptor B2 primary antibodies
  • Description
    Rabbit polyclonal to Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y604)
  • Host species
    Rabbit
  • Specificity
    ab61791 detects endogenous levels of EPHB1/2 only when phosphorylated at tyrosine 594 and tyrosine 604.
  • Tested applications
    Suitable for: WB, ICC/IF, ELISAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human Eph receptor B1 + Eph receptor B2.

  • Positive control
    • Extracts from HepG2 cells, HuvEc cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab61791 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
WB 1/500 - 1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa).
ICC/IF 1/500 - 1/1000.
ELISA 1/5000.

Target

  • Relevance
    Ephrin receptors bind members of the ephrin ligand family. They are divided into 2 groups based on the similarity of their extracellular domain sequences and their affinities for binding ephrin-A and ephrin-B ligands. They make up the largest subgroup of the receptor tyrosine kinase (RTK) family. Eph receptor B1 (EphB1) and Eph receptor B2 (EphB2) are receptors for ephrin-B family members. Developing and adult neural tissue express nearly all of the Ephrin receptors and ephrin ligands. Ephrins and ephrin receptors also play a significant role in angiogenesis. EphB2 acts as a tumor suppressor.
  • Cellular localization
    Membrane; Single-pass type I membrane protein.
  • Database links
  • Alternative names
    • EPH B1 antibody
    • EPH B2 antibody
    • Eph tyrosine kinase 2 antibody
    • Eph tyrosine kinase 3 antibody
    • EPHB1 antibody
    • EPHB2 antibody
    • Ephrin receptor EphB1 antibody
    • Ephrin receptor EphB2 antibody
    • Ephrin type B receptor 1 antibody
    • Ephrin type B receptor 2 antibody
    • EPHT 2 antibody
    • EPHT 3 antibody
    • ERK antibody
    • Hek 5 antibody
    • Hek 6 antibody
    • Tyrosine protein kinase receptor EPH2 antibody
    • Tyrosine protein kinase receptor EPH3 antibody
    see all

Images

  • All lanes : Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y604) antibody (ab61791) at 1/1000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HeLa whole cell lysate - overexpressing EphB2
    Lane 3 : HeLa whole cell lysate - overexpressinf EphB2-kinase dead (K661R)

    Lysates/proteins at 100000 cells per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG monoclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 110 kDa
    Observed band size: 110 kDa


    Exposure time: 5 minutes


    All cells were stimulated with preclustered ephrinb1-Fc chimeric protein.

    See Abreview

  • ab61791 staining Eph receptor B1 + Eph receptor B2 (phospho Y594) in HeLa cells (overexpressing  EphB2 and stimulated for 20 minutes with preclustered efnb1-Fc chimeric protein) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS + 1% BSA + 0.1% Triton X-100) for 2 hour at 25°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody. Red - anti-EphB2 (goat), Grey - DAPI, Green - ab61791.

    See Abreview

  • All lanes : Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y604) antibody (ab61791) at 1/500 dilution

    Lane 1 : Extracts from HepG2 cells
    Lane 2 : Extracts from HepG2 cells with the immunising phosphopeptide

    Predicted band size: 110 kDa
    Observed band size: 110 kDa

  • Immunofluorescence analysis of HuvEc cells, using ab61791 at 1/500-1/1000 dilution with (+) or without (-) immunising phosphopeptide.

References

This product has been referenced in:
  • Mimche PN  et al. EphB2 receptor tyrosine kinase promotes hepatic fibrogenesis in mice via activation of hepatic stellate cells. Sci Rep 8:2532 (2018). IHC-Fr . Read more (PubMed: 29416088) »
  • Clements MP  et al. The Wound Microenvironment Reprograms Schwann Cells to Invasive Mesenchymal-like Cells to Drive Peripheral Nerve Regeneration. Neuron 96:98-114.e7 (2017). WB . Read more (PubMed: 28957681) »
See all 8 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Loading amount
100000 cells
Gel Running Conditions
Reduced Denaturing (4-20%)
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Treatment
stimulated with 1´g/ml clustered ephrinb1-Fc
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Thomas Jungas

Verified customer

Submitted Apr 11 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Human Cell (HeLa cells)
Specification
HeLa cells
Permeabilization
Yes - Triton X-100 @ 0.1%
Fixative
Paraformaldehyde

Mr. Thomas Jungas

Verified customer

Submitted Mar 21 2014

Answer

ab61791 detects Eph B1 receptor phosphorylation at Y594 and the corresponding phosphorylation status of Eph B2 receptor at Y596.

In terms of the significance of Eph B2 receptor phosphorylation at Y596, a literature search through some of the references listed in the phosphosite webpage for Y596 below may be of help:
http://www.phosphosite.org/siteAction.do?id=3532#1

I also performed a quick literature search and found the article below, which mentions:
"Alternatively, phosphorylation of Y596/604might aid or induce receptor oligomerization, resulting intrans phosphorylation of Y779. Experiments to order the phosphorylation events and to determine whether juxtamembrane phosphorylation is a prerequisite for YACT-based activation are under way."
http://mcb.asm.org/content/20/13/4791.full

Since this article is from 2000, it may be worth contacting the authors to see if they have further information regarding how the activation status of Eph B2 is related to phosphorylation of Y596.

Read More

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