Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Frog - Xenopus (African Clawed Frog), Transgenic Mouse

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunofluorescence, Immunocytochemistry, Immunoprecipitation, Functional Assay, Proximity Ligation Assay (PLA)

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Ephrin-B2 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Ephrin-B2
Arg27-Ala227
Accession # AAA82934

Specificity

Detects mouse Ephrin-B2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 5% cross-reactivity with recombinant mouse Ephrin-B1 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat Ephrin‑B2 Antibody

Detection of Mouse Ephrin‑B2 antibody by Western Blot.

Detection of Mouse Ephrin‑B2 by Western Blot.

Western blot shows lysates of mouse embryo tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat Ephrin-B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF496) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF019). Specific bands were detected for Ephrin-B2 at approximately 40-45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Detection of Ephrin-B2 antibody in SH-SY5Y Human Cell Line antibody by Flow Cytometry.

Detection of Ephrin‑B2 in SH‑SY5Y Human Cell Line by Flow Cytometry.

SH-SY5Y human neuroblastoma cell line was stained with Goat Anti-Human/Mouse/Rat Ephrin-B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF496, filled histogram) or isotype control antibody (AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (F0107).
Ephrin-B2 antibody in Rat Hippo-campal Neurons by Immunocytochemistry (ICC).

Ephrin‑B2 in Rat Hippo-campal Neurons.

Ephrin‑B2 was detected in immersion fixed rat hippocampal neurons using 2 µg/mL Goat Anti-Human/Mouse/Rat Ephrin‑B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF496) for 3 hours at room temperature. Cells were stained (red) and counterstained (green). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Ephrin‑B2 in SH‑SY5Y Human Cell Line.

Ephrin‑B2 was detected in immersion fixed SH‑SY5Y human neuroblastoma cell line using Goat Anti-Human/Mouse/Rat Ephrin‑B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # af496) at 15 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to the cell surface. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Ephrin-B2 by Western Blot

Detection of Ephrin-B2 by Western Blot

Effects of Hsp90 inhibitors on LANA in KSHV-infected endothelial cells.(A) L1T2 cells were treated with AUY922 at concentrations of 0, 0.02, 0.1, 0.5, and 2.5 µM for 48 hours, whole cell lysates were immunoblotted with anti-LANA, anti-Hsp90 antibodies, anti-EphA2, anti-Ephrin-B2, and anti-Akt (total), and anti-pAkt (S473) antibodies separately. Apoptosis was evaluated with anti-cleaved PARP and anti-cleaved Caspase-3 antibodies, Cdc2 and beta -Actin were used as controls. (B–C) SLK-KSHV were treated with 0.5 µM 17-DMAG for 0, 12, 24, 48, 72 and 96 hours, or at concentrations of 0, 0.1, 0.5, 2.5 and 10 µM for 48 hours separately, whole cell lysates were immunoblotted with anti-LANA antibody, anti-EphA2, anti-EphrinB2, Cdc2 and beta -Actin were used as controls. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23209418), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Ephrin-B2 by Immunohistochemistry

Detection of Human Ephrin-B2 by Immunohistochemistry

Gene signatures peculiar to the venular compartment. (A) EFNB2 expression measured by scRNA-seq (left) and immunofluorescence staining (right) denoting its abundance across different BEC subpopulations and its exclusive expression in human dermal BVs (white arrowheads). (B) Representative confocal images of skin sections (upper) and whole-mount skin blocks (lower) stained for ERG (green), EFNB2 (red) and ICAM1 (white). The unique composition of EFNB2 and ICAM1 expression enables the discrimination of arterioles (empty arrowheads), post-capillary venules (*) and venules (white arrowheads). (C) EGR2 expression is most enriched in the post-capillary venule cluster and is restricted to BVs (white arrowheads). (D) Representative section (upper) and whole-mount staining (lower) of ERG (green), EGR2 (red) and ICAM1 (white), highlighting EGR2+ICAM1+ BVs. Scale bars: 100 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35406678), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Ephrin-B2 by Immunohistochemistry

Detection of Human Ephrin-B2 by Immunohistochemistry

Gene signatures peculiar to the venular compartment. (A) EFNB2 expression measured by scRNA-seq (left) and immunofluorescence staining (right) denoting its abundance across different BEC subpopulations and its exclusive expression in human dermal BVs (white arrowheads). (B) Representative confocal images of skin sections (upper) and whole-mount skin blocks (lower) stained for ERG (green), EFNB2 (red) and ICAM1 (white). The unique composition of EFNB2 and ICAM1 expression enables the discrimination of arterioles (empty arrowheads), post-capillary venules (*) and venules (white arrowheads). (C) EGR2 expression is most enriched in the post-capillary venule cluster and is restricted to BVs (white arrowheads). (D) Representative section (upper) and whole-mount staining (lower) of ERG (green), EGR2 (red) and ICAM1 (white), highlighting EGR2+ICAM1+ BVs. Scale bars: 100 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35406678), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat Ephrin‑B2 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: SH‑SY5Y human neuroblastoma cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed cultured rat embryonic hippocampal neurons and fixed SH-SY5Y human neuroblastoma cell line

Western Blot

1 µg/mL
Sample: Mouse embryo tissue

Reviewed Applications

Read 3 reviews rated 4.3 using AF496 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Ephrin-B2

Ephrin-B2, also known as Htk-L, ELF-2, LERK-5, and NLERK-1 (1), is a member of the ephrin ligand family which binds members of the Eph receptor family. All ligands share a conserved extracellular sequence, which most likely corresponds to the receptor binding domain. This conserved sequence consists of approximately 125 amino acids and includes four invariant cysteines. The B-class ligands are transmembrane proteins which can become tyrosine phosphorylated upon receptor ligation. The cytoplasmic domains are approximately 80 amino acids long and are highly conserved, especially the last 33 amino acids. Several signaling molecules have been shown to interact with the cytoplasmic region, although specific signaling roles have yet to be elucidated. Ephrin-B2 has been shown to bind EphA4, EphB1, EphB2, EphB3, and EphB4 (2, 3). The extracellular domains of murine and human Ephrin-B2 share 98% amino acid identity. Only membrane-bound or Fc-clustered ligands are capable of activating the receptor in vitro. While soluble monomeric ligands bind the receptor, they do not induce receptor autophosphorylation and activation (2). In vivo, the ligands and receptors display reciprocal expression (3). It has been found that nearly all the receptors and ligands are expressed in developing and adult neural tissue (3). The Ephrin/Eph families also appear to play a role in angiogenesis (3).

References

  1. Eph Nomenclature Committee [letter]. (1997) Cell 90:403.
  2. Flanagan, J.G. and P. Vanderhaeghen (1998) Annu. Rev. Neurosci. 21:309.
  3. Pasquale, E.B. (1997) Curr. Opin. Cell Biol. 9:608.

Alternate Names

EFNB2, ELF-2, EphrinB2, Htk-L, LERK-5, NLERK-1

Entrez Gene IDs

1948 (Human); 13642 (Mouse); 30219 (Zebrafish)

Gene Symbol

EFNB2

UniProt

AAA82934

Additional Ephrin-B2 Products

Product Documents for Human/Mouse/Rat Ephrin‑B2 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat Ephrin‑B2 Antibody

For research use only

Citations for Human/Mouse/Rat Ephrin‑B2 Antibody

Customer Reviews for Human/Mouse/Rat Ephrin‑B2 Antibody (3)

4.3 out of 5
3 Customer Ratings
5 Stars
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4 Stars
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3 Stars
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Human/Mouse/Rat Ephrin-B2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cell Lysates and Cell culture supernatant
    Species: Human
    Verified Customer | Posted 03/16/2021
    Human/Mouse/Rat Ephrin‑B2 Antibody AF496
  • Human/Mouse/Rat Ephrin-B2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: E12.5 mouse embryo fixed in 4% PFA
    Species: Mouse
    Verified Customer | Posted 12/16/2020
    Dilution used - 1:200. The staining was done on an E12.5 transverse mouse section fixed overnight with 4% PFA. Stained using standard IF technique. Attached picture shows an artery and a vein and it is clearly seen that the artery seen is stained positive whereas the adjacent vein is negative for EphrinB2 (as it should be).
    Human/Mouse/Rat Ephrin‑B2 Antibody AF496
  • Human/Mouse/Rat Ephrin-B2 Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: mouse brain lysate
    Species: Mouse
    Verified Customer | Posted 07/12/2017
    Simple Western lane view shows 53 and 58kDa signals for ephrinB2. Note that higher concentrations of this antibody may lead to an unspecific signal corresponding to the size of the internal standard (230 kDa, leftmost lane).
    Simple Western: anti-ephrinB2 antibody [biotechne AF496] Simple Western lane view shows 53 and 58kDa signals for ephrinB2 in 0.5 mg/ml of mouse brain lysate (lysis buffer from ARY014 proteome profiler kit). This run was performed under reducing conditions using the 12-230 kDa separation system in combination with a secondary anti-goat-HRP antibody (biotechne, HAF109, 1:50). Note that higher concentrations of this antibody may lead to an unspecific signal corresponding to the size of the internal standard (230 kDa, leftmost lane).
    Human/Mouse/Rat Ephrin‑B2 Antibody AF496

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