Human/Mouse EphB2 Antibody

(23 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • 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.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse EphB2
    Val27-Lys548
    Accession # P54763
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Simple Western
    20 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunohistochemistry
    1-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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 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 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.
Immunocytochemistry
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.
Immunohistochemistry
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.
Immunohistochemistry
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.
Immunohistochemistry
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 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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
  • Entrez Gene IDs:
    2048 (Human); 13844 (Mouse)
  • Alternate Names:
    CAPB; Cek5; Drt; DRTEphB2; EC 2.7.10; EC 2.7.10.1; EK5; elk-related tyrosine kinase; EPH receptor B2; eph tyrosine kinase 3; EphB2; EPH-like kinase 5; ephrin type-B receptor 2; EPHT3MGC87492; EPTH3; Erk; ERKHek5; Hek5; Nuk; PCBC; protein-tyrosine kinase HEK5; Qek2; Renal carcinoma antigen NY-REN-47; Sek3; Tyro5; Tyrosine-protein kinase receptor EPH-3; Tyrosine-protein kinase TYRO5
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

23 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon
    Authors: YH Liu, JW Tsai, JL Chen, WS Yang, PC Chang, PL Cheng, DL Turner, Y Yanagawa, TW Wang, JY Yu
    Sci Rep, 2017;7(0):42895.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  2. Generation of Mouse and Human Organoid-Forming Intestinal Progenitor Cells by Direct Lineage Reprogramming
    Authors: S Miura, A Suzuki
    Cell Stem Cell, 2017;0(0):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC
  3. Lipid-mediated Wnt protein stabilization enables serum-free culture of human organ stem cells
    Authors: N Tüysüz, L van Bloois, S van den Br, H Begthel, MM Verstegen, LJ Cruz, L Hui, LJ van der La, J de Jonge, R Vries, E Braakman, E Mastrobatt, JJ Cornelisse, H Clevers, D Ten Berge
    Nat Commun, 2017;8(0):14578.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  4. Induction of endoplasmic reticulum stress by deletion of Grp78 depletes Apc mutant intestinal epithelial stem cells
    Oncogene, 2016;0(0):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC - Paraffin embedded
  5. EphrinB2 repression through ZEB2 mediates tumour invasion and anti-angiogenic resistance
    Nat Commun, 2016;7(0):12329.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  6. The transcription factor XBP1s restores hippocampal synaptic plasticity and memory by control of the Kalirin-7 pathway in Alzheimer model
    Mol Psychiatry, 2016;0(0):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Frozen
  7. GluA2 trafficking is involved in apoptosis of retinal ganglion cells induced by activation of EphB/EphrinB reverse signaling in a rat chronic ocular hypertension model.
    Authors: Dong L, Gao F, Wang X, Miao Y, Wang S, Wu Y, Li F, Wu J, Cheng X, Sun X, Yang X, Wang Z
    J Neurosci, 2015;35(13):5409-21.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  8. Tissue-Specific Effects of Reduced beta-catenin Expression on Adenomatous Polyposis Coli Mutation-Instigated Tumorigenesis in Mouse Colon and Ovarian Epithelium.
    Authors: Feng Y, Sakamoto N, Wu R, Liu J, Wiese A, Green M, Green M, Akyol A, Roy B, Zhai Y, Cho K, Fearon E
    PLoS Genet, 2015;11(11):e1005638.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC - paraffin
  9. Instruction of circulating endothelial progenitors in vitro towards specialized blood-brain barrier and arterial phenotypes.
    Authors: Boyer-Di Ponio J, El-Ayoubi F, Glacial F, Ganeshamoorthy K, Driancourt C, Godet M, Perriere N, Guillevic O, Couraud P, Uzan G
    PLoS ONE, 2014;9(1):e84179.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC Fluorescence
  10. A mathematical model for eph/ephrin-directed segregation of intermingled cells.
    Authors: Aharon, Rotem, Janes, Peter W, Burgess, Anthony, Hamza, Kais, Klebaner, Fima, Lackmann, Martin
    PLoS ONE, 2014;9(12):e111803.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  11. Reelin induces EphB activation.
    Authors: Bouche E, Romero-Ortega M, Henkemeyer M, Catchpole T, Leemhuis J, Frotscher M, May P, Herz J, Bock H
    Cell Res, 2013;23(4):473-90.
  12. Smad3 contributes to positioning of proliferating cells in colonic crypts by inducing EphB receptor protein expression.
    Authors: Furukawa K, Sato T, Katsuno T, Nakagawa T, Noguchi Y, Tokumasa A, Yokote K, Yokosuka O, Saito Y
    Biochem. Biophys. Res. Commun., 2011;405(4):521-6.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  13. Neuropsin cleaves EphB2 in the amygdala to control anxiety.
    Authors: Attwood BK, Bourgognon JM, Patel S, Mucha M, Schiavon E, Skrzypiec AE, Young KW, Shiosaka S, Korostynski M, Piechota M, Przewlocki R, Pawlak R
    Nature, 2011;473(7347):372-5.
    Species: Mouse
    Sample Type: In Vivo
    Application: In vivo
  14. Ephrin-B1 regulates axon guidance by reverse signaling through a PDZ-dependent mechanism.
    Authors: Bush JO, Soriano P
    Genes Dev., 2009;23(13):1586-99.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: IP
  15. Ephrin-B reverse signaling promotes structural and functional synaptic maturation in vivo.
    Authors: Lim BK, Matsuda N, Poo MM
    Nat. Neurosci., 2008;11(2):160-9.
    Species: Xenopus
    Sample Type: Tissue Homogenates
    Application: IP
  16. EphrinB2 regulation by PTH and PTHrP revealed by molecular profiling in differentiating osteoblasts.
    Authors: Allan EH, Hausler KD, Wei T, Gooi JH, Quinn JM, Crimeen-Irwin B, Pompolo S, Sims NA, Gillespie MT, Onyia JE, Martin TJ
    J. Bone Miner. Res., 2008;23(8):1170-81.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  17. Fish oil increases cholesterol storage in white adipose tissue with concomitant decreases in inflammation, hepatic steatosis, and atherosclerosis in mice.
    Authors: Saraswathi V, Gao L, Morrow JD, Chait A, Niswender KD, Hasty AH
    J. Nutr., 2007;137(7):1776-82.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  18. Phosphorylation of ephrin-B1 regulates dissemination of gastric scirrhous carcinoma.
    Authors: Tanaka M, Kamata R, Takigahira M, Yanagihara K, Sakai R
    Am. J. Pathol., 2007;171(1):68-78.
    Species: Mouse
    Sample Type: Whole Cells
    Application: WB
  19. Cupredoxin-cancer interrelationship: azurin binding with EphB2, interference in EphB2 tyrosine phosphorylation, and inhibition of cancer growth.
    Authors: Chaudhari A, Mahfouz M, Fialho AM, Yamada T, Granja AT, Zhu Y, Hashimoto W, Schlarb-Ridley B, Cho W, Das Gupta TK, Chakrabarty AM
    Biochemistry, 2007;46(7):1799-810.
    Species: Human
    Sample Type: Cell Lysates
    Application: IP
  20. EphB receptors coordinate migration and proliferation in the intestinal stem cell niche.
    Authors: Holmberg J, Genander M, Halford MM, Anneren C, Sondell M, Chumley MJ, Silvany RE, Henkemeyer M, Frisen J
    Cell, 2006;125(6):1151-63.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: IP
  21. EphB2 and ephrin-B1 expressed in the adult kidney regulate the cytoarchitecture of medullary tubule cells through Rho family GTPases.
    Authors: Ogawa K, Wada H, Okada N, Harada I, Nakajima T, Pasquale EB, Tsuyama S
    J. Cell. Sci., 2006;119(0):559-70.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC
  22. EphB2/R-Ras signaling regulates glioma cell adhesion, growth, and invasion.
    Authors: Nakada M, Niska JA, Tran NL, McDonough WS, Berens ME
    Am. J. Pathol., 2005;167(2):565-76.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  23. The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells.
    Authors: Nakada M, Niska JA, Miyamori H, McDonough WS, Wu J, Sato H, Berens ME
    Cancer Res., 2004;64(9):3179-85.
    Species: Human
    Sample Type: Cell Lysates
    Application: IP
Expand to show all 23 Citations
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Isotype Controls
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Average Rating: 4 (Based on 1 review)

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We have 1 review tested in 1 application: Western Blot.

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Western Blot Human/Mouse EphB2 Antibody AF467
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Very Good
 WB Mouse Anonymous 11/10/2017
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Western Blot Human/Mouse EphB2 Antibody AF467
Western Blot: Human/Mouse EphB2 Antibody [AF467]

Summary

ApplicationWestern Blot
Sample TestedLung tissue
SpeciesMouse

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