AF3038

antibody from R&D Systems

Targeting: EPHB4 HTK, Tyro11

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

AF3038

Provider

R&D Systems

Product name

Human EphB4 Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human EphB4 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse (rm) EphB4 is observed and less than 1% cross-reactivity with recombinant human (rh) EphA4, rhEphA3, rhEphB2, and rhEphB3 is observed.

Reactivity

Human

Host

Goat

Conjugate

Unconjugated

Antigen sequence

P54760


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

Downregulation of proinflammatory cytokines in HTLV-1-infected T cells by Resveratrol.

Fuggetta MP, Bordignon V, Cottarelli A, Macchi B, Frezza C, Cordiali-Fei P, Ensoli F, Ciafrè S, Marino-Merlo F, Mastino A, Ravagnan G
Journal of experimental & clinical cancer research : CR 2016 Jul 22;35(1):118

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EPHB4 kinase-inactivating mutations cause autosomal dominant lymphatic-related hydrops fetalis.

Martin-Almedina S, Martinez-Corral I, Holdhus R, Vicente A, Fotiou E, Lin S, Petersen K, Simpson MA, Hoischen A, Gilissen C, Jeffery H, Atton G, Karapouliou C, Brice G, Gordon K, Wiseman JW, Wedin M, Rockson SG, Jeffery S, Mortimer PS, Snyder MP, Berland S, Mansour S, Makinen T, Ostergaard P
The Journal of clinical investigation 2016 Aug 1;126(8):3080-8

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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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BCR-ABL-independent and RAS / MAPK pathway-dependent form of imatinib resistance in Ph-positive acute lymphoblastic leukemia cell line with activation of EphB4.

Suzuki M, Abe A, Imagama S, Nomura Y, Tanizaki R, Minami Y, Hayakawa F, Ito Y, Katsumi A, Yamamoto K, Emi N, Kiyoi H, Naoe T
European journal of haematology 2010 Mar;84(3):229-38

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Soluble EphB4 inhibition of PDGF-induced RPE migration in vitro.

He S, Kumar SR, Zhou P, Krasnoperov V, Ryan SJ, Gill PS, Hinton DR
Investigative ophthalmology & visual science 2010 Jan;51(1):543-52

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Activation of the receptor EphB4 by its specific ligand ephrin B2 in human osteoarthritic subchondral bone osteoblasts.

Kwan Tat S, Pelletier JP, Amiable N, Boileau C, Lajeunesse D, Duval N, Martel-Pelletier J
Arthritis and rheumatism 2008 Dec;58(12):3820-30

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human EphB4 by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, COLO 205 human colorectal adenocarcinoma cell line, ZR-75 human breast cancer cell line, and HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EphB4 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human EphB4 by Simple Western^TM. Simple Western lane view shows lysates of HUVEC human umbilical vein endothelial cells, loaded at 0.2 mg/mL. A specific band was detected for EphB4 at approximately 127 kDa (as indicated) using 50 µg/mL of Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038) 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.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Western Blot Shows Human EphB4 Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and EphB4 knockout HEK293T cell line (KO). PVDF membrane was probed with 2 µg/mL of Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EphB4 at approximately 140 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

EphB4 in Human Kidney. EphB4 was detected in immersion fixed paraffin-embedded sections of human kidney using 15 µg/mL Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038) overnight at 4 °C. Tissue was stained with 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 Paraffin-embedded Tissue Sections.

Flow cytometry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of EphB4 in MCF-7 Human Cell Line by Flow Cytometry. MCF-7 human breast cancer cell line was stained with Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038, 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.