EPHB4 antibody | AbD01327

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Human anti EPHB4

Product Type
Monoclonal Antibody
Clone
AbD01327
Isotype
HuCAL Fab bivalent
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
HCA001 E P WB 0.1 mg
Human anti EPHB4, clone AbD01327 recognizes the receptor tyrosine kinase EPHB4, a member of the largest known family of these kinases, and plays an important role in a variety of processes during embryonic development including pattern formation, cell aggregation and migration, segmentation, neural development, angiogenesis, and vascular network assembly.

EPHB4, along with its ligand, Ephrin-B2, is essential for vascular remodeling, maturation and directed growth. Both Ephrin-B2 and EPHB4 targeted gene knock-outs in mice show lack of remodeling of the primitive vascular plexus to form mature vascular structures. EPHB4 expression is restricted to venous endothelial cells. EPHB4 and Ephrin-B2 are also essential for defining the boundaries between arterial and venous domains, which persist in the adult. EPHB4 selectively binds Ephrin-B2 and not other Ephrin B ligands.

The B class Ephrins are comprised of transmembrane proteins with an intracellular domain that can elaborate reverse signaling. Ligand-receptor binding leads to protein clustering followed by receptor activation. Furthermore, interaction between EPHB4 and Ephrin-B2 can provoke bidirectional signaling. It has been shown in vitro that EPHB4-mediated forward signaling restricts intermingling of cells and supports cellular segregation, whereas reverse signaling from Ephrin-B2 stimulates migration and sprouting angiogenesis. Downstream signaling by EPHBs regulates cell proliferation, cytoskeletal organization, and migration.

EPHB4 is abundantly expressed in placenta and in a range of primary tissues and malignant cell lines. Expressed in foetal, but not adult, brain, and in primitive and myeloid, but not lymphoid, hematopoietic cells.

In Western blotting, clone AbD01327 may cross-react with EPHB1, however in immunohistology the staining pattern is consistent with specificity only for EPHB4.

Product Details

Target Species
Human
Species Cross-Reactivity
Target SpeciesCross Reactivity
Mouse Expected from Sequence
N.B. Antibody reactivity and working conditions may vary between species.
Product Form
A bivalent human recombinant Fab selected from the HuCAL® GOLD phage display library. Expressed in E. coli and purified using NiNTA affinity chromatography. This Fab fragment is dimerized via a helix-turn-helix motif. The antibody is tagged with a myc-tag (EQKLISEEDL) and a his-tag (HHHHHH) at the C-terminus of the anti-body heavy chain. This antibody is supplied lyophilised.
Reconstitution
Reconstitute with 1.0ml distilled water
Preparation
Metal chelate affinity chromatography
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
1% Bovine Serum Albumin
Immunogen
Recombinant EPHB4 fusion protein (Leu563 - Tyr987), corresponding to the intracellular part of the protein, including the kinase domain and an SAM (sterile alpha motif) domain.
Approx. Protein Concentrations
0.1 mg/ml after reconstitution

Storage Information

Storage
Prior to reconstitution store at +4oC.
After reconstitution store at -20oC.
Storage in frost-free freezers is not recommended. This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
6 months from date of reconstitution.

More Information

UniProt
P54760 Related reagents
Entrez Gene
EPHB4 Related reagents
GO Terms
GO:0005003 ephrin receptor activity
GO:0007169 transmembrane receptor protein tyrosine kinase signaling pathway
GO:0005887 integral to plasma membrane
GO:0005524 ATP binding
GO:0006468 protein phosphorylation
GO:0009887 organ morphogenesis
GO:0008283 cell proliferation
GO:0009986 cell surface
GO:0045765 regulation of angiogenesis
Acknowledgements
Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
His-tag is a registered trademark of EMD Biosciences.
Licensed Use
For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
Regulatory
For research purposes only

Applications of EPHB4 antibody

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
ELISA 2ug/ml
Immunohistology - Paraffin 10ug/ml
Western Blotting 2ug/ml
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Technical Advice
Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
ELISA
Due to the presence of bovine serum albumin (BSA), this antibody is unsuitable for use as a capture reagent in sandwich ELISA applications. This product is also available without BSA, please enquire.

Secondary Antibodies Available

Description Product Code Pack Size Applications List Price Quantity
Goat anti Human IgG F(ab')2:HRP 0500-0099 0.5 ml E WB
Goat anti Human IgG F(ab')2:Alk. Phos. 0500-0100 0.5 ml E WB
Mouse anti c-Myc:HRP MCA2200P 0.1 mg C E P WB*
Mouse anti Penta Histidine Tag:HRP MCA5995P 125 µl E WB
Goat anti Human IgG F(ab')2:Alk. Phos STAR126A 0.5 ml E WB
Goat anti Human IgG F(ab')2:Biotin STAR126B 0.5 mg C E F WB
Goat anti Human IgG F(ab')2:HRP STAR126P 0.5 ml E WB

Product Specific References

References for EPHB4 antibody

  1. Xia, G. et al. (2005) EphB4 expression and biological significance in prostate cancer.
    Cancer Res. 65: 4623-4632.