HCA001 recognises 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.
Specifically, 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, HCA001 may cross-react with EphB1, however in immunohistology the staining pattern is consistent with specificity only for EPHB4.
Species Cross-Reactivity
Target Species
Cross Reactivity
Mouse
Expected from Sequence
Product Form
A bivalent human recombinant Fab selected from the HuCAL® GOLD phage display library. Expressed inE. coliand purified using NiNTA affinity chromatography. This Fab fragment is dimerized via a helix-turn-helix mo-tif. 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.
Application
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 visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
2ug/ml
Flow Cytometry
Immunohistology - Frozen
Immunohistology - Paraffin
10ug/ml
Immunoprecipitation
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.
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.
Licensed Use
For in vitro research purposes only, unless otherwise specified in writing by AbD Serotec.
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.
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