EPHB4 Antibody | 3934

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Staining of formalin fixed paraffin embedded human placenta tissue with HCA025, using goat anti human alkaline phosphatase secondary antibody (AHP1108P)

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  • Human anti EPHB4
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  • Product Type
    Monoclonal Antibody
  • Clone
    3934
  • Isotype
    HuCAL Fab bivalent
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    HCA025E, P *, WBdatasheet pdfdatasheet pdf0.1 mg
    HCA025
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
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    • Human anti EPHB4 antibody recognizes the receptor tyrosine kinase EPHB4. EPHB4 is 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, Human anti EPHB4 antibody may cross-react with EPHB1, however in immunohistology the staining pattern is consistent with specificity only for EPHB4
    • Intended Use
    • Target Species
      Human
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      MouseExpected from Sequence
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      A lyophilised, 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 bivalent by dimerization of the bacterial alkaline phosphatase fusion protein. The antibody is tagged with a DYKDDDDK tag and a Strep tag at the C-terminus of the antibody heavy chain.
    • Reconstitution
      Reconstitute with 0.25ml distilled water
    • Preparation
      Metal chelate affinity chromatography
    • Preservative Stabilisers
      None present
    • 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
    • Purity
    • Approx. Protein Concentrations
      0.4mg/ml after reconstitution
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Storage
      Prior to reconstitution store at +4oC.
      After reconstitution store at -20oC.
      Storage in frost-free freezers is not recommended. 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.
    • GO Terms
      organ morphogenesis
      regulation of angiogenesis
      transmembrane receptor protein tyrosine kinase signaling pathway
      integral to plasma membrane
      protein phosphorylation
      ATP binding
      ephrin receptor activity
      cell proliferation
      cell surface
    • UniProt
    • Entrez Gene
    • Acknowledgements
      Sold under license of U.S. Patents 6,300,064, 6,696,248, 6,708,484, 6,753,136, European Patent 0,859,841 and corresponding patents.
    • Licensed Use
      For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
    • Regulatory
      For research purposes only
    • 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 NameYesNoMin DilutionMax Dilution
      ELISA2ug/ml
      Western Blotting2ug/ml
      Immunohistology - Paraffin(1)10ug/ml
      (1)
      This product requires antigen retrieval using heat treatment prior to staining of paraffin sections.Sodium citrate buffer pH 6.0 is recommended for this purpose.

    • 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
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional EPHB4 Antibody Formats

    Formats Clone Applications Sizes available
    EPHB4 Antibody : Purified 3934 E, P *, WB 0.1 mg
    • Copyright © 2016 Bio-Rad

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      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              References

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