Anti-Ephrin B2 antibody (ab96264)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab96264 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 36 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionCell surface transmembrane ligand for Eph receptors, a family of receptor tyrosine kinases which are crucial for migration, repulsion and adhesion during neuronal, vascular and epithelial development. Binds promiscuously Eph receptors residing on adjacent cells, leading to contact-dependent bidirectional signaling into neighboring cells. The signaling pathway downstream of the receptor is referred to as forward signaling while the signaling pathway downstream of the ephrin ligand is referred to as reverse signaling. Binds to receptor tyrosine kinase including EPHA4, EPHA3 and EPHB4. Together with EPHB4 plays a central role in heart morphogenesis and angiogenesis through regulation of cell adhesion and cell migration. EPHB4-mediated forward signaling controls cellular repulsion and segregation from EFNB2-expressing cells. May play a role in constraining the orientation of longitudinally projecting axons.
  • Tissue specificityLung and kidney.
  • Sequence similaritiesBelongs to the ephrin family.
    Contains 1 ephrin RBD (ephrin receptor-binding) domain.
  • Post-translational
    modifications
    Inducible phosphorylation of tyrosine residues in the cytoplasmic domain.
  • Cellular localizationMembrane.
  • Target information above from: UniProt accession P52799 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • EFN B2 antibody
    • EFNB 2 antibody
    • EFNB2 antibody
    • EFNB2_HUMAN antibody
    • Eph related receptor tyrosine kinase ligand 5 antibody
    • EPH-related receptor tyrosine kinase ligand 5 antibody
    • ephrin B2 antibody
    • Ephrin-B2 antibody
    • EphrinB2 antibody
    • EPLG 5 antibody
    • EPLG5 antibody
    • Htk L antibody
    • HTK ligand antibody
    • HTK-L antibody
    • HTKL antibody
    • LERK 5 antibody
    • LERK-5 antibody
    • LERK5 antibody
    • Ligand of eph related kinase 5 antibody
    • MGC126226 antibody
    • MGC126227 antibody
    • MGC126228 antibody
    • OTTMUSP00000024973 antibody
    see all

Anti-Ephrin B2 antibody images

  • All lanes : Anti-Ephrin B2 antibody (ab96264) at 1 µg/ml

    Lane 1 : Lung (Human) Tissue Lysate
    Lane 2 : Kidney (Human) Tissue Lysate - adult normal tissue (ab30203)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 32 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 73 kDa,86 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
  • ICC/IF image of ab96264 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab96264, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Ephrin B2 antibody (ab96264)

ab96264 has not yet been referenced specifically in any publications.

Product Wall

Thank you for your reply.

I am happy to hear the staining has improved!

The antibody was raised against an extracellular sequence, but since it is a polyclonal it really could have multiple binding epitopes throughout the protein an...

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Thank you for your reply.

Yes, I think you should include the blocking buffer in with the secondary. Typically we recommend 1% BSA, but 5% BSA should be okay.

http://www.abcam.com/index.html?pageconfig=resource&rid=14733
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Thank you for taking time to contact us. I am sorry to hear that this antibody is not providing satisfactory results.

So the antibody is staining well in immunocytochemistry (ICC/IF)on fixed human endothelial cells? What is the fixative you'...

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Let me just confirm it is the products Anti-Ephrin B2 antibody (ab96264) and Anti-Ephrin B2 (phospho Y316) antibody (ab119323) which you are interested in?

The anti-Ephrin B2 antibody (ab96264) does indeed measure the total Ephrin B2 content,...

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Thank you for calling Abcam earlier today.

I am sorry about the problems you were having with ab96264 in IF. As we discussed, I am sending you a new vial of the antibody to test. Your new order/reference number is *******.

If there ...

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