AF467

antibody from R&D Systems

Targeting: EPHB2 DRT, EPHT3, ERK, Hek5, Tyro5

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

AF467

Provider

R&D Systems

Product name

Human/Mouse EphB2 Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. 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.

Reactivity

Human, Mouse

Host

Goat

Conjugate

Unconjugated

Antigen sequence

P54763


Isotype

IgG

Vial size

100 ug

Concentration

LYOPH

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.

References

Submitted references

EphB2-dependent signaling promotes neuronal excitotoxicity and inflammation in the acute phase of ischemic stroke.

Ernst AS, Böhler LI, Hagenston AM, Hoffmann A, Heiland S, Sticht C, Bendszus M, Hecker M, Bading H, Marti HH, Korff T, Kunze R
Acta neuropathologica communications 2019 Feb 5;7(1):15

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EphB2 receptor tyrosine kinase promotes hepatic fibrogenesis in mice via activation of hepatic stellate cells.

Mimche PN, Lee CM, Mimche SM, Thapa M, Grakoui A, Henkemeyer M, Lamb TJ
Scientific reports 2018 Feb 7;8(1):2532

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Generation of Mouse and Human Organoid-Forming Intestinal Progenitor Cells by Direct Lineage Reprogramming.

Miura S, Suzuki A
Cell stem cell 2017 Oct 5;21(4):456-471.e5

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The transcription factor XBP1s restores hippocampal synaptic plasticity and memory by control of the Kalirin-7 pathway in Alzheimer model.

Cissé M, Duplan E, Lorivel T, Dunys J, Bauer C, Meckler X, Gerakis Y, Lauritzen I, Checler F
Molecular psychiatry 2017 Nov;22(11):1562-1575

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Ascl1 promotes tangential migration and confines migratory routes by induction of Ephb2 in the telencephalon.

Liu YH, Tsai JW, Chen JL, Yang WS, Chang PC, Cheng PL, Turner DL, Yanagawa Y, Wang TW, Yu JY
Scientific reports 2017 Mar 9;7:42895

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Lipid-mediated Wnt protein stabilization enables serum-free culture of human organ stem cells.

Tüysüz N, van Bloois L, van den Brink S, Begthel H, Verstegen MM, Cruz LJ, Hui L, van der Laan LJ, de Jonge J, Vries R, Braakman E, Mastrobattista E, Cornelissen JJ, Clevers H, Ten Berge D
Nature communications 2017 Mar 6;8:14578

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Induction of endoplasmic reticulum stress by deletion of Grp78 depletes Apc mutant intestinal epithelial stem cells.

van Lidth de Jeude JF, Meijer BJ, Wielenga MCB, Spaan CN, Baan B, Rosekrans SL, Meisner S, Shen YH, Lee AS, Paton JC, Paton AW, Muncan V, van den Brink GR, Heijmans J
Oncogene 2017 Jun 15;36(24):3397-3405

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Unidirectional Eph/ephrin signaling creates a cortical actomyosin differential to drive cell segregation.

O'Neill AK, Kindberg AA, Niethamer TK, Larson AR, Ho HH, Greenberg ME, Bush JO
The Journal of cell biology 2016 Oct 24;215(2):217-229

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Blocking the Interaction between EphB2 and ADDLs by a Small Peptide Rescues Impaired Synaptic Plasticity and Memory Deficits in a Mouse Model of Alzheimer's Disease.

Shi XD, Sun K, Hu R, Liu XY, Hu QM, Sun XY, Yao B, Sun N, Hao JR, Wei P, Han Y, Gao C
The Journal of neuroscience : the official journal of the Society for Neuroscience 2016 Nov 23;36(47):11959-11973

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EphrinB2 repression through ZEB2 mediates tumour invasion and anti-angiogenic resistance.

Depner C, Zum Buttel H, Böğürcü N, Cuesta AM, Aburto MR, Seidel S, Finkelmeier F, Foss F, Hofmann J, Kaulich K, Barbus S, Segarra M, Reifenberger G, Garvalov BK, Acker T, Acker-Palmer A
Nature communications 2016 Jul 29;7:12329

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Tissue-Specific Effects of Reduced β-catenin Expression on Adenomatous Polyposis Coli Mutation-Instigated Tumorigenesis in Mouse Colon and Ovarian Epithelium.

Feng Y, Sakamoto N, Wu R, Liu JY, Wiese A, Green ME, Green M, Akyol A, Roy BC, Zhai Y, Cho KR, Fearon ER
PLoS genetics 2015 Nov;11(11):e1005638

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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.

Dong LD, Gao F, Wang XH, Miao Y, Wang SY, Wu Y, Li F, Wu J, Cheng XL, Sun XH, Yang XL, Wang Z
The Journal of neuroscience : the official journal of the Society for Neuroscience 2015 Apr 1;35(13):5409-21

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A mathematical model for eph/ephrin-directed segregation of intermingled cells.

Aharon R, Janes PW, Burgess AW, Hamza K, Klebaner F, Lackmann M
PloS one 2014;9(12):e111803

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Instruction of circulating endothelial progenitors in vitro towards specialized blood-brain barrier and arterial phenotypes.

Boyer-Di Ponio J, El-Ayoubi F, Glacial F, Ganeshamoorthy K, Driancourt C, Godet M, Perrière N, Guillevic O, Couraud PO, Uzan G
PloS one 2014;9(1):e84179

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Reelin induces EphB activation.

Bouché E, Romero-Ortega MI, Henkemeyer M, Catchpole T, Leemhuis J, Frotscher M, May P, Herz J, Bock HH
Cell research 2013 Apr;23(4):473-90

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Neuropsin cleaves EphB2 in the amygdala to control anxiety.

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 May 19;473(7347):372-5

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Smad3 contributes to positioning of proliferating cells in colonic crypts by inducing EphB receptor protein expression.

Furukawa K, Sato T, Katsuno T, Nakagawa T, Noguchi Y, Tokumasa A, Yokote K, Yokosuka O, Saito Y
Biochemical and biophysical research communications 2011 Feb 25;405(4):521-6

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Ephrin-B1 regulates axon guidance by reverse signaling through a PDZ-dependent mechanism.

Bush JO, Soriano P
Genes & development 2009 Jul 1;23(13):1586-99

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Ephrin-B reverse signaling promotes structural and functional synaptic maturation in vivo.

Lim BK, Matsuda N, Poo MM
Nature neuroscience 2008 Feb;11(2):160-9

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EphrinB2 regulation by PTH and PTHrP revealed by molecular profiling in differentiating osteoblasts.

Allan EH, Häusler KD, Wei T, Gooi JH, Quinn JM, Crimeen-Irwin B, Pompolo S, Sims NA, Gillespie MT, Onyia JE, Martin TJ
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 2008 Aug;23(8):1170-81

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Phosphorylation of ephrin-B1 regulates dissemination of gastric scirrhous carcinoma.

Tanaka M, Kamata R, Takigahira M, Yanagihara K, Sakai R
The American journal of pathology 2007 Jul;171(1):68-78

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Fish oil increases cholesterol storage in white adipose tissue with concomitant decreases in inflammation, hepatic steatosis, and atherosclerosis in mice.

Saraswathi V, Gao L, Morrow JD, Chait A, Niswender KD, Hasty AH
The Journal of nutrition 2007 Jul;137(7):1776-82

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Cupredoxin-cancer interrelationship: azurin binding with EphB2, interference in EphB2 tyrosine phosphorylation, and inhibition of cancer growth.

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 Feb 20;46(7):1799-810

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EphB receptors coordinate migration and proliferation in the intestinal stem cell niche.

Holmberg J, Genander M, Halford MM, Annerén C, Sondell M, Chumley MJ, Silvany RE, Henkemeyer M, Frisén J
Cell 2006 Jun 16;125(6):1151-63

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EphB2 and ephrin-B1 expressed in the adult kidney regulate the cytoarchitecture of medullary tubule cells through Rho family GTPases.

Ogawa K, Wada H, Okada N, Harada I, Nakajima T, Pasquale EB, Tsuyama S
Journal of cell science 2006 Feb 1;119(Pt 3):559-70

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EphB2/R-Ras signaling regulates glioma cell adhesion, growth, and invasion.

Nakada M, Niska JA, Tran NL, McDonough WS, Berens ME
The American journal of pathology 2005 Aug;167(2):565-76

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The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells.

Nakada M, Niska JA, Miyamori H, McDonough WS, Wu J, Sato H, Berens ME
Cancer research 2004 May 1;64(9):3179-85

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The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells.

Nakada M, Niska JA, Miyamori H, McDonough WS, Wu J, Sato H, Berens ME
Cancer research 2004 May 1;64(9):3179-85

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human EphB2 by Simple Western^TM. 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.

Immunocytochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Flow cytometry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.