Invitrogen

EphB4 Monoclonal Antibody (3D7G8)

Product Image Gallery

EphB4 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of EphB4 (green) in HT29 cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a EphB4 mouse monoclonal antibody (Product # 37-1800) at a concentration of 5 µg/mL in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731) at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

EphB4 Antibody in Immunohistochemistry (IHC) — Immunohistochemical staining of human colon carcinoma tissue using EphB4 Receptor Monoclonal Antibody, Mouse (clone 3D7G8) (clone 3D7G8) (Product # 37-1800).

EphB4 Antibody in Western Blot (WB) — Western blot analysis of EphB4 was performed by loading 20 µg of indicated whole cell lysate and 7 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. EphB4 was detected at ~125 kDa using a EphB4 mouse monoclonal antibody (Product # 37-1800, upper panel) at a concentration of 2 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177) at a dilution of 1:2000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076). GAPDH was detected at ~37 kDa using a GAPDH rabbit polyclonal antibody, HRP conjugate (Product # PA1-987-HRP, lower panel).

EphB4 Antibody — Figure 5 .Immunohistochemistry was used to detect ephrin-B and EphB4 protein expression. Representative immunostaining of a (A) negative ephrin-B tumor, (B) membranous ephrin-B, (C) EphB4-negative, (D) nuclear EphB4 and (E) membranous EphB4. Scale bars, 100 u m. (F) EphB4 antibody validation in MCF7 cells was performed by immunoblotting and the intensity of the bands quantified (n=2). Error bars represent the standard error of the mean (SEM). The AllStar siRNA was used as negative control and GAPDH siRNA as positive control. The ephrin B antibody was the same used in all the in vitro experiments. {KD}

EphB4 Antibody in Immunocytochemistry, Immunohistochemistry (Paraffin), Flow Cytometry (ICC/IF, IHC (P), Flow) — Figure 3 Presence of EphA2 and EphB4 on colon cancer cells. ( A ) Flow cytometry of HT-29 shows abundant presence of both EphA2 and EphB4 compared with negative Jurkat cells; ( B ) Immunofluorescence of EphB4 indicates a membranous staining on HT-29 colon cancer cells grown in monolayer. X-axes indicate fluorescence intensity, the scale bar represents 20 micrometers.

EphB4 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Figure 1 Two examples of staining patterns for EphA2 and EphB4 in sets of tumor and normal tissue from patients with colon cancer. ( A ) shows the most generally found pattern, with low expression of both proteins in normal tissue and abundant expression in tumor tissue; ( B ) shows an aberrant expression pattern (as indicated in Figure 2 ), with extremely high EphA2 expression in normal tissue and absence of staining in the corresponding tumor tissue. The scale bars represent 150 micrometers; ( C ) shows 40x enlargements of the sections indicated in A. Red arrows indicate membranous staining. The scale bar represents 30 micrometer.

EphB4 Antibody in Peptide array (ARRAY) — Figure 2 N/T scorings diagram for the tissue microarray (TMA) stained for EphA2 and EphB4. Green = higher score in tumor than normal tissue; Red = higher score in normal tissue than tumor tissue; Blue = no difference in score between normal and tumor tissue. The thickness of the line represents the number of patients, the thicker the line the more patients. The numbers 0-7 represent the IHC scoring values.

Product Details

37-1800

Applications	Tested Dilution	Publications
Western Blot (WB)	1-2 µg/mL	View 3 publications 3 publications
Immunohistochemistry (IHC)	Assay-dependent	View 2 publications 2 publications
Immunohistochemistry (Paraffin) (IHC (P))	-	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	3-5 µg/mL	View 2 publications 2 publications
Flow Cytometry (Flow)	-	View 1 publication 1 publication
ELISA (ELISA)	Assay-dependent	-
Immunoprecipitation (IP)	Assay-dependent	View 1 publication 1 publication
Peptide Array (Array)	-	View 1 publication 1 publication
Miscellaneous PubMed (Misc)	-	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human
Published species	Hamster, Human, Mouse
Host/Isotype	Mouse / IgG1, kappa
Class	Monoclonal
Type	Antibody
Clone	3D7G8
Immunogen	Recombinant protein derived from the C-terminal region of the human EphB4 protein. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.4
Contains	0.1% sodium azide
Storage conditions	-20°C
RRID	AB_2533303

Product Specific Information

37-1800 has been successfully used in Western Blot, immunohistochemical, immunofluorescence, ELISA and immunoprecipitation assays. By western blot this antibody detects a ~125kDa protein representing EPHB4 in MCF7, HT-29 and U2OS lysate.

Target Information

Ephrin receptors and their ligands, the ephrins, mediate numerous developmental processes, particularly in the nervous system. Based on their structures and sequence relationships, ephrins are divided into the ephrin-A (EFNA) class, which are anchored to the membrane by a glycosylphosphatidylinositol linkage, and the ephrin-B (EFNB) class, which are transmembrane proteins. The Eph family of receptors are divided into 2 groups based on the similarity of their extracellular domain sequences and their affinities for binding ephrin-A and ephrin-B ligands. Ephrin receptors make up the largest subgroup of the receptor tyrosine kinase (RTK) family. The protein encoded by this gene binds to ephrin-B2 and plays an essential role in vascular development.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Evaluation of EphB4 as Target for Image-Guided Surgery of Breast Cancer.

Pharmaceuticals (Basel, Switzerland)

de Muijnck C,van Gorkom Y,van Duijvenvoorde M,Eghtesadi M,Dekker-Ensink G,Bhairosingh SS,Affinito A,Kuppen PJK,Vahrmeijer AL,Sier CFM

37-1800 was used in Immunohistochemistry to prove that EphB4 is a promising candidate for targeted image-guided surgery of breast cancer, especially for Her2 negative cases.

Thu Jul 30 00:00:00 EDT 2020

Evaluation of EphB4 as Target for Image-Guided Surgery of Breast Cancer.

Pharmaceuticals (Basel, Switzerland)

de Muijnck C,van Gorkom Y,van Duijvenvoorde M,Eghtesadi M,Dekker-Ensink G,Bhairosingh SS,Affinito A,Kuppen PJK,Vahrmeijer AL,Sier CFM

37-1800 was used in Immunohistochemistry to prove that EphB4 is a promising candidate for targeted image-guided surgery of breast cancer, especially for Her2 negative cases.

Thu Jul 30 00:00:00 EDT 2020

Ephrin‑B2 inhibits cell proliferation and motility in vitro and predicts longer metastasis‑free survival in breast cancer.

International journal of oncology

Magic Z,Sandström J,Perez-Tenorio G

Published figure using EphB4 monoclonal antibody (Product # 37-1800) in Western Blot

Sun Dec 01 00:00:00 EST 2019

Human blood vessel organoids as a model of diabetic vasculopathy.

Nature

Wimmer RA,Leopoldi A,Aichinger M,Wick N,Hantusch B,Novatchkova M,Taubenschmid J,Hämmerle M,Esk C,Bagley JA,Lindenhofer D,Chen G,Boehm M,Agu CA,Yang F,Fu B,Zuber J,Knoblich JA,Kerjaschki D,Penninger JM

37-1800 was used in Immunocytochemistry-immunoflourescence to suggest that blood vessel organoids derived from human stem cells faithfully recapitulate the structure and function of human blood vessels and are a suitable model for diabetic vasculopathy.

Tue Jan 01 00:00:00 EST 2019

Evaluation of EphA2 and EphB4 as Targets for Image-Guided Colorectal Cancer Surgery.

International journal of molecular sciences

Stammes MA,Prevoo HA,Ter Horst MC,Groot SA,Van de Velde CJ,Chan AB,de Geus-Oei LF,Kuppen PJ,Vahrmeijer AL,Pasquale EB,Sier CF

371800 was used in immunohistochemistry - paraffin section to examine if EphA2 and EphB4 proteins are suitable targets for image-guided oncologic surgery of colorectal cancer

Fri Feb 03 00:00:00 EST 2017

Evaluation of EphA2 and EphB4 as Targets for Image-Guided Colorectal Cancer Surgery.

International journal of molecular sciences

Stammes MA,Prevoo HA,Ter Horst MC,Groot SA,Van de Velde CJ,Chan AB,de Geus-Oei LF,Kuppen PJ,Vahrmeijer AL,Pasquale EB,Sier CF

371800 was used in immunohistochemistry - paraffin section to examine if EphA2 and EphB4 proteins are suitable targets for image-guided oncologic surgery of colorectal cancer

Fri Feb 03 00:00:00 EST 2017

Evaluation of EphA2 and EphB4 as Targets for Image-Guided Colorectal Cancer Surgery.

International journal of molecular sciences

Stammes MA,Prevoo HA,Ter Horst MC,Groot SA,Van de Velde CJ,Chan AB,de Geus-Oei LF,Kuppen PJ,Vahrmeijer AL,Pasquale EB,Sier CF

371800 was used in immunohistochemistry - paraffin section to examine if EphA2 and EphB4 proteins are suitable targets for image-guided oncologic surgery of colorectal cancer

Fri Feb 03 00:00:00 EST 2017

Evaluation of EphA2 and EphB4 as Targets for Image-Guided Colorectal Cancer Surgery.

International journal of molecular sciences

Stammes MA,Prevoo HA,Ter Horst MC,Groot SA,Van de Velde CJ,Chan AB,de Geus-Oei LF,Kuppen PJ,Vahrmeijer AL,Pasquale EB,Sier CF

371800 was used in immunohistochemistry - paraffin section to examine if EphA2 and EphB4 proteins are suitable targets for image-guided oncologic surgery of colorectal cancer

Fri Feb 03 00:00:00 EST 2017

Erythropoietin Stimulates Tumor Growth via EphB4.

Cancer cell

Pradeep S,Huang J,Mora EM,Nick AM,Cho MS,Wu SY,Noh K,Pecot CV,Rupaimoole R,Stein MA,Brock S,Wen Y,Xiong C,Gharpure K,Hansen JM,Nagaraja AS,Previs RA,Vivas-Mejia P,Han HD,Hu W,Mangala LS,Zand B,Stagg LJ,Ladbury JE,Ozpolat B,Alpay SN,Nishimura M,Stone RL,Matsuo K,Armaiz-Peña GN,Dalton HJ,Danes C,Goodman B,Rodriguez-Aguayo C,Kruger C,Schneider A,Haghpeykar S,Jaladurgam P,Hung MC,Coleman RL,Liu J,Li C,Urbauer D,Lopez-Berestein G,Jackson DB,Sood AK

Published figure using EphB4 monoclonal antibody (Product # 37-1800) in Immunohistochemistry

Mon Nov 09 00:00:00 EST 2015

YES oncogenic activity is specified by its SH4 domain and regulates RAS/MAPK signaling in colon carcinoma cells.

American journal of cancer research

Dubois F,Leroy C,Simon V,Benistant C,Roche S

37-1800 was used in immunoprecipitation to elucidate how YES contributes to colorectal cancer progression

Thu Aug 13 00:00:00 EDT 2015

Gene expression profiling identifies EPHB4 as a potential predictive biomarker in colorectal cancer patients treated with bevacizumab.

Medical oncology (Northwood, London, England)

Guijarro-Muñoz I,Sánchez A,Martínez-Martínez E,García JM,Salas C,Provencio M,Alvarez-Vallina L,Sanz L

37-1800 was used in immunohistochemistry (paraffin) to investigate the prognostic value of EPHB4 in colorectal cancer therapy.

Fri Oct 25 00:00:00 EDT 2013

EPHB4 and survival of colorectal cancer patients.

Cancer research

Davalos V,Dopeso H,Castaño J,Wilson AJ,Vilardell F,Romero-Gimenez J,Espín E,Armengol M,Capella G,Mariadason JM,Aaltonen LA,Schwartz S,Arango D

37-1800 was used in immunohistochemistry - paraffin section and western blot to study the role of EPHB4 in colorectal cancer and determine its value as a prognostic marker

Fri Sep 15 00:00:00 EDT 2006

EphB receptor-binding peptides identified by phage display enable design of an antagonist with ephrin-like affinity.

The Journal of biological chemistry

Koolpe M,Burgess R,Dail M,Pasquale EB

Fri Apr 29 00:00:00 EDT 2005

EphB receptor-binding peptides identified by phage display enable design of an antagonist with ephrin-like affinity.

The Journal of biological chemistry

Koolpe M,Burgess R,Dail M,Pasquale EB

Fri Apr 29 00:00:00 EDT 2005

EphB receptor-binding peptides identified by phage display enable design of an antagonist with ephrin-like affinity.

The Journal of biological chemistry

Koolpe M,Burgess R,Dail M,Pasquale EB

Fri Apr 29 00:00:00 EDT 2005

EphB receptor-binding peptides identified by phage display enable design of an antagonist with ephrin-like affinity.

The Journal of biological chemistry

Koolpe M,Burgess R,Dail M,Pasquale EB

Fri Apr 29 00:00:00 EDT 2005

Bioinformatics

Protein Aliases: CAPB; DRT; EPHB2; EPHB3; Ephrin B1; Ephrin B2; Ephrin B3; Ephrin B4; ephrin receptor EphB4; Ephrin type-B receptor 4; EphrinB1; EphrinB2; EphrinB3; EphrinB4; EPHT3; ERK; ETK2; HEK-6; HEK2; Hek5; Hepatoma transmembrane kinase; MGC87492; PCBC; soluble EPHB4 variant 1; soluble EPHB4 variant 2; soluble EPHB4 variant 3; Tyro5; TYRO6; tyrosine-protein kinase receptor HTK; Tyrosine-protein kinase TYRO11

Gene Aliases: EPHB4; HTK; MYK1; TYRO11

UniProt ID: (Human) P54760

Entrez Gene ID: (Human) 2050

Molecular Function: transmembrane signal receptor