Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

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

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunofluorescence, Immunocytochemistry, Simple Western, Immunoprecipitation, Proximity Ligation Assay, In vivo assay, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all EphB2 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse EphB2
Val27-Lys548
Accession # P54763

Specificity

Detects mouse and human EphB2 in direct ELISAs and Western blots. In Western blots, approximately 5% cross-reactivity with recombinant rat (rr) EphB1, recombinant mouse (rm) EphA8, rmEphA6, rmEphB6, rmEphA3, rmEphA4, rmEphA7, rrEphA5, recombinant human EphA1, rmEphA2 and rmEphB3 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse EphB2 Antibody

Detection of Recombinant Human and Mouse EphB2 antibody by Western Blot.

Detection of Recombinant Human and Mouse EphB2 by Western Blot.

Western blot shows 25 ng of Recombinant Human EphB2 Fc Chimera (Catalog # 5189-B2) and Recombinant Mouse EphB2 Fc Chimera (Catalog # 467-B2). PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for EphB2 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Detection of EphB2 antibody in COLO 205 Human Cell Line antibody by Flow Cytometry.

Detection of EphB2 in COLO 205 Human Cell Line by Flow Cytometry.

COLO 205 human colorectal adenocarcinoma cell line was stained with Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Detection of EphB2 antibody in D3 Mouse Cell Line antibody by Flow Cytometry.

Detection of EphB2 in D3 Mouse Cell Line by Flow Cytometry.

D3 mouse embryonic stem cell line was stained with Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

EphB2 antibody in MBA-MB-468 Human Cell Line by Immunocytochemistry (ICC).

EphB2 in MBA‑MB‑468 Human Cell Line.

EphB2 was detected in immersion fixed MBA-MB-468 human breast cancer cell line using Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467) at 5 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 cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

EphB2 antibody in Embryonic Mouse Brain by Immunohistochemistry (IHC-Fr).

EphB2 in Embryonic Mouse Brain.

EphB2 was detected in immersion fixed frozen sections of embryonic mouse brain (15 d.p.c.) using 15 µg/mL Goat Anti-Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467) overnight at 4 °C. Tissue was stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained (green). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

EphB2 antibody in Human Esophageal Squamous Cell Carcinoma by Immunohistochemistry (IHC-P).

EphB2 in Human Esophageal Squamous Cell Carcinoma.

EphB2 was detected in immersion fixed paraffin-embedded sections of human esophageal squamous cell carcinoma using Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

EphB2 antibody in Mouse Embryo by Immunohistochemistry (IHC-Fr).

EphB2 in Mouse Embryo.

EphB2 was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c.) using Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to developing brain. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human EphB2 by Simple WesternTM.

Simple Western lane view shows lysates of Exosome Standards (COLO1) (NBP2-49845) and COLO 205 human colorectal adenocarcinoma cell line, loaded at 0.5 mg/ml. A specific band was detected for EphB2 at approximately 135 kDa (as indicated) using 1 µg/ml of Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467) followed by HRP-conjugated Donkey Anti-Goat Secondary Antibody (Catalog # 042-206). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Detection of Human EphB2 antibody by Simple WesternTM.

Detection of Human EphB2 by Simple WesternTM.

Simple Western lane view shows lysates of COLO 205 human colorectal adenocarcinoma cell line and MBA-MB-468 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for EphB2 at approximately 139 and 146 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse EphB2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF467) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human EphB2 by Immunocytochemistry/Immunofluorescence

Detection of Human EphB2 by Immunocytochemistry/Immunofluorescence

Colorectal tumours maintain tissue organisation similar to normal colon.Detection of EPHB2 (green) and ERBB3 (red, A and B) by co-immunofluorescence in normal colon (A) and colorectal cancer (B) (DAPI, blue). Scale bar, 50μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26367378), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse EphB2 by Immunocytochemistry/Immunofluorescence

Detection of Mouse EphB2 by Immunocytochemistry/Immunofluorescence

Expression of EphB2 increases and is activated on HSCs after chronic CCl4 -induced liver injury. (a) Isolated cell fractions from livers of mice subjected to chronic CCl4 injections were analyzed for EphB2, Ephrin-B1, Ephrin-B2 and Ephrin-B3 mRNA levels using RT-qPCR. Results are shown as fold change compared to liver cell fractions obtained from vehicle-treated controls. Error bars represent mean ± SEM.; n = 6 animals; CD11b = macrophages, LSEC = Liver sinusoidal endothelial cells, HEP = Hepatocytes and HSCs = Hepatic stellate cells. (b) OCT liver sections from C57BL/6 J mice chronically injected with CCl4 or vehicle (oil) controls were stained with EphB2 (red), alpha SMA (green) and DAPI/DNA (blue) and analyzed using confocal microscopy. Scale bar = 100 µm, “C” denotes the central vein. All images are representative of 5 mice per group. (c) OCT liver sections from C57BL/6 J mice chronically injected with CCl4 or vehicle controls were stained with phospho-EphB1/EphB2-Y594 (red), PDGFR beta (green) and DAPI/DNA (blue) and analyzed using confocal microscopy. Scale bar = 50 µm. All images are representative of 5 mice per group. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29416088), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human EphB2 by Western Blot

Detection of Human EphB2 by Western Blot

Increased cell-cell adhesion within one cell population is required for the formation of tightly packed cell clusters.A) Simulation of cell-cell segregation using the same adhesion term in both cell types (Aeph  =  Aephrin  = 100, left panel) vs. increased adhesion only in the green cell population (Aeph  = 110, Aephrin  = 100, right panel). Both simulations started with the same number of Eph (green) and ephrin (black) expressing cells. In the “Equal adhesion” case, an ‘Islands-in-a-sea’ pattern is less apparent. B) Representative images from segregation assays of unlabelled ephrin-B1 cells co-cultured with Cell Tracker-green labelled (green staining) EphB2 cells, without (left) or with E-cadherin-cherry expression (red staining, right); scale bar, 75 µm. C) Quantitation of cell densities in the cell clusters shown in B (n = 10). D) Western blot analysis of lysates from parental and E-cadherin-cherry-transduced cells, using the indicated antibodies. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0111803), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse EphB2 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: COLO 205 human colorectal adenocarcinoma cell line and D3 mouse embryonic stem cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed MDA-MB-468 human breast cancer cell line

Immunohistochemistry

1-15 µg/mL
Sample: Immersion fixed frozen sections of embryonic mouse brain (15 d.p.c.), immersion fixed paraffin-embedded sections of human esophageal squamous cell carcinoma, and immersion fixed frozen sections of mouse embryo (13 d.p.c.)

Simple Western

1-20 µg/mL
Sample: Exosome Standards (COLO1) (Catalog # NBP2-49845), COLO 205 human colorectal adenocarcinoma cell line and MBA‑MB‑468 human breast cancer cell line

Western Blot

0.1 µg/mL
Sample: Recombinant Human EphB2 Fc Chimera (Catalog # 5189-B2) and Recombinant Mouse EphB2 Fc Chimera (Catalog # 467-B2).

Reviewed Applications

Read 9 reviews rated 4.3 using AF467 in the following applications:

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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: EphB2

EphB2, also known as Cek5, Nuk, Erk, Qek2, Tyro5, Sek3, Hek5, and Drt (1), is a member of the Eph receptor family which binds members of the ephrin ligand family. There are two classes of receptors, designated A and B. Both the A and B class receptors have an extracellular region consisting of a globular domain, a cysteine-rich domain, and two fibronectin type III domains. This is followed by the transmembrane region and the cytoplasmic region. The cytoplasmic region contains a juxtamembrane motif with two tyrosine residues which are the major autophosphorylation sites, a kinase domain, and a conserved sterile alpha motif (SAM) in the carboxy tail which contains one conserved tyrosine residue. Activation of kinase activity occurs after ligand recognition and binding. EphB2 has been shown to bind ephrin-B1, ephrin-B2, and ephrin-B3 (2, 3). The extracellular domains of human and mouse EphB2 share 99% amino acid identity. Only membrane-bound or
Fc‑clustered ligands are capable of activating the receptor in vitro. Soluble monomeric ligands bind the receptor but 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.

Long Name

Eph Receptor B2

Alternate Names

Cek5, Drt, Erk, Hek5, Nuk, Qek2, Sek3, Tyro5

Entrez Gene IDs

2048 (Human); 13844 (Mouse)

Gene Symbol

EPHB2

UniProt

P54763

Additional EphB2 Products

Product Documents for Human/Mouse EphB2 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse EphB2 Antibody

For research use only

Citations for Human/Mouse EphB2 Antibody

Customer Reviews for Human/Mouse EphB2 Antibody (9)

4.3 out of 5
9 Customer Ratings
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Customer Images

Showing  1 - 5 of 9 reviews Showing All
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  • Human/Mouse EphB2 Antibody
    Name: STAVROULA THEOFANOUS
    Application: Western Blot
    Sample Tested: brain and spinal cord
    Species: Mouse
    Verified Customer | Posted 07/05/2018
    Lane1: Precision Plus Protein™ Dual Color Standards7ul Lane2:Ephb1 receptor (SC tissue from naive C57BL6j mouse)
    Blot Loading • ANTIBODY: Human/Mouse EphB2Antibody cat noAF542 Lane1: Precision Plus Protein™ Dual Color Standards7ul Lane2:Ephb1 receptor (SC tissue from naive C57BL6j mouse) Western blot experiment details Gel:4-20% SDS page Tranfer apparatus: Trans-Blot® Turbo™ Transfer System Membrane: midi size nitrocellulose Time: 25min at 75-125V Primary ab:Goat –rat EphB1 Ab 1mg for 20h Secondary ab: rabbit a-goat 1:5,000 for 1h Substrate: HRP for 15min
    Human/Mouse EphB2 Antibody AF467
  • Human/Mouse EphB2 Antibody
    Name: STAVROULA THEOFANOUS
    Application: Immunoprecipitation
    Sample Tested: brain and spinal cord
    Species: Mouse
    Verified Customer | Posted 04/16/2018
    Blot Loading • ANTIBODY: Human/Mouse EphB2Antibody cat noAF467 Lane1: PreĐision Plus Protein™ Dual Color Standards8ul Lane2:Ephb2 receptors IP 1mg protein (SC tissue) Lane3:Ephb2 receptors no IP 25ug (SC tissue) Lane4:Goat GFP IP (negative control instead of goat IgG)1mg (SC tissue) Lane5:Ephb2 receptors IP 1mg (SC tissue) Lane6:Ephb2 receptors no IP 25ug (SC tissue) Lane7:Goat GFP IP (negative control instead of goat IgG)1mg (SC tissue) Western blot experiment details Gel:4-20% SDS page Tranfer apparatus: Trans-Blot® Turďo™ Transfer System Membrane: midi size nitrocellulose Time: 25min at 75-125V Primary ab:Goat –m/h EphB2 Ab 1mg for 20h Secondary ab: donkey a-goat 1:15,000 for 1h Substrate: HRP for 15min
    Human/Mouse EphB2 Antibody AF467
  • Human/Mouse EphB2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Mouse iPS
    Species: Mouse
    Verified Customer | Posted 01/17/2018
    Human/Mouse EphB2 Antibody AF467
  • Human/Mouse EphB2 Antibody
    Name: Li Liu
    Application: Simple Western
    Sample Tested: Adult lung
    Species: Mouse
    Verified Customer | Posted 01/12/2018
  • Human/Mouse EphB2 Antibody
    Name: Dharmaraja Allimuthu
    Application: Western Blot
    Sample Tested: Embryonic stem cells
    Species: Human
    Verified Customer | Posted 12/21/2017
    Human/Mouse EphB2 Antibody AF467
  • Human/Mouse EphB2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Lung tissue
    Species: Mouse
    Verified Customer | Posted 11/10/2017
    Human/Mouse EphB2 Antibody AF467
  • Human/Mouse EphB2 Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: mouse brain lysate
    Species: Mouse
    Verified Customer | Posted 07/12/2017
    Simple Western: anti-EphB2 antibody [biotechne AF467] Simple Western lane view shows a specific 128 kDa signal for EphB2 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).
    Human/Mouse EphB2 Antibody AF467
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 21276420
    Species: Mouse
    Verified Customer | Posted 01/08/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22144690
    Species: Other
    Verified Customer | Posted 01/08/2015

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