Ephrin-B1 Antibody [Unconjugated]

Western blot shows lysates of mouse lung tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse Ephrin‑B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF473) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Ephrin‑B1 at approximately 45-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Ephrin-B1 was detected in immersion fixed frozen sections of mouse kidney using Goat Anti-Mouse Ephrin-B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF473) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Simple Western lane view shows lysates of mouse lung tissue and mouse spleen tissue, loaded at 0.2 mg/mL. A specific band was detected for Ephrin‑B1 at approximately 65 kDa (as indicated) using 25 µg/mL of Goat Anti-Mouse Ephrin‑B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF473) 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.

• Reactivity: Mu
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Ephrin-B1 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Ephrin-B1
  Lys30-Ser229
  Accession # AAA53231
• Specificity: Detects mouse Ephrin-B1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) Ephrin-A3, rhEphrin-A4, recombinant mouse (rm) Ephrin-A1, rmEphrin-B2, rhEphrin-A5, rhEphrin-B3, rmEphrin-A2, rmEphrin‑A4, and recombinant zebrafish Ephrin‑B2 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: Efnb1

Applications/Dilutions

• Dilutions:
  • Western Blot 0.25 ug/mL
  • Simple Western 25 ug/mL
  • Immunohistochemistry 5-15 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Ephrin-B1 Antibody [Unconjugated]

• Cek5-L
• EFL3
• EFL-3
• EFNB1
• ELK ligand
• ELK-L
• EphrinB1
• Ephrin-B1
• LERK-2
• STRA-1

Background

Ephrin-B1, also known as LERK-2, ELK-L, EFL-3, Cek5-L, and STRA-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-B1 has been shown to bind EphA3, EphB1, EphB2, EphB3, and EphB4 (2, 3). The extracellular domains of human and mouse Ephrin-B1 share 94% 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 receptors and ligands are expressed in developing and adult neural tissue (3). The Eph/ephrin families also appear to play a role in angiogenesis (3).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Ephrin-B1 Antibody (AF473)(15)

Publications using AF473

• J Koeppen, AQ Nguyen, AM Nikolakopo, M Garcia, S Hanna, S Woodruff, Z Figueroa, A Obenaus, IM Ethell Functional consequences of synapse remodeling following astrocyte-specific regulation of ephrin-B1 in the adult hippocampus J. Neurosci., 2018;0(0):. 2018 [PMID: 29793972] (IHC-P, Mouse)
• AK O'Neill, AA Kindberg, TK Niethamer, AR Larson, HH Ho, ME Greenberg, JO Bush Unidirectional Eph/ephrin signaling creates a cortical actomyosin differential to drive cell segregation J. Cell Biol., 2016;215(2):217-229. 2016 [PMID: 27810913] (IHC, Mouse)
• Geng X, Cha B, Mahamud M, Lim K, Silasi-Mansat R, Uddin M, Miura N, Xia L, Simon A, Engel J, Chen H, Lupu F, Srinivasan R Multiple mouse models of primary lymphedema exhibit distinct defects in lymphovenous valve development. Dev Biol, 2016;409(1):218-33. 2016 [PMID: 26542011] (IHC, Mouse)
• Feng Y, Sakamoto N, Wu R, Liu J, Wiese A, Green M, Green M, Akyol A, Roy B, Zhai Y, Cho K, Fearon E Tissue-Specific Effects of Reduced beta-catenin Expression on Adenomatous Polyposis Coli Mutation-Instigated Tumorigenesis in Mouse Colon and Ovarian Epithelium. PLoS Genet, 2015;11(11):e1005638. 2015 [PMID: 26528816] (IHC-P, Mouse)
• Dong L, Gao F, Wang X, Miao Y, Wang S, Wu Y, Li F, Wu J, Cheng X, Sun X, Yang X, Wang Z 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. J Neurosci, 2015;35(13):5409-21. 2015 [PMID: 25834064] (Western Blot, Rat)
• Arvanitis D, Behar A, Drougard A, Roullet P, Davy A Cortical abnormalities and non-spatial learning deficits in a mouse model of CranioFrontoNasal syndrome. PLoS ONE, 2014;9(2):e88325. 2014 [PMID: 24520368] (IHC, Mouse)
• Dimidschstein J, Passante L, Dufour A, van den Ameele J, Tiberi L, Hrechdakian T, Adams R, Klein R, Lie D, Jossin Y, Vanderhaeghen P Ephrin-B1 controls the columnar distribution of cortical pyramidal neurons by restricting their tangential migration. Neuron, 2013;79(6):1123-35. 2013 [PMID: 24050402] (Immunoprecipitation, Mouse)
• Wu Z, Luo H, Thorin E, Tremblay J, Peng J, Lavoie JL, Wang Y, Qi S, Wu T, Wu J Possible role of Efnb1 protein, a ligand of Eph receptor tyrosine kinases, in modulating blood pressure. J. Biol. Chem., 2012;287(19):15557-69. 2012 [PMID: 22393061] (Western Blot, Mouse)
• Lim BK, Matsuda N, Poo MM Ephrin-B reverse signaling promotes structural and functional synaptic maturation in vivo. Nat. Neurosci., 2008;11(2):160-9. 2008 [PMID: 18193042] (Immunoprecipitation, Xenopus)
• Dietrich T, Onderka J, Bock F, Kruse FE, Vossmeyer D, Stragies R, Zahn G, Cursiefen C Inhibition of inflammatory lymphangiogenesis by integrin alpha5 blockade. Am. J. Pathol., 2007;171(1):361-72. 2007 [PMID: 17591980] (ICC, Mouse)
• Gupta S, Ramjaun AR, Haiko P, Wang Y, Warne PH, Nicke B, Nye E, Stamp G, Alitalo K, Downward J Binding of ras to phosphoinositide 3-kinase p110alpha is required for ras-driven tumorigenesis in mice. Cell, 2007;129(5):957-68. 2007 [PMID: 17540175] (IHC-Fr, Mouse)
• Kayser MS, McClelland AC, Hughes EG, Dalva MB Intracellular and trans-synaptic regulation of glutamatergic synaptogenesis by EphB receptors. J. Neurosci., 2006;26(47):12152-64. 2006 [PMID: 17122040] (ICC, Rat)
• Yang NY, Pasquale EB, Owen LB, Ethell IM The EphB4 receptor-tyrosine kinase promotes the migration of melanoma cells through Rho-mediated actin cytoskeleton reorganization. J. Biol. Chem., 2006;281(43):32574-86. 2006 [PMID: 16950769] (Western Blot, Mouse)
• Ogawa K, Wada H, Okada N, Harada I, Nakajima T, Pasquale EB, Tsuyama S EphB2 and ephrin-B1 expressed in the adult kidney regulate the cytoarchitecture of medullary tubule cells through Rho family GTPases. J. Cell. Sci., 2006;119(0):559-70. 2006 [PMID: 16443753] (IHC, Mouse)
• Yu G, Luo H, Wu Y, Wu J EphrinB1 is essential in T-cell-T-cell co-operation during T-cell activation. J. Biol. Chem., 2004;279(53):55531-9. 2004 [PMID: 15502157] (Flow Cytometry, Mouse)

Bioinformatics

• Gene Symbol: Efnb1
• Uniprot:
  • AAA53231()