Invitrogen

EphB2 Monoclonal Antibody (1A6C9)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated. View Details

8 Published Figures

12 References

View all (24) EphB2 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

EphB2 Antibody in Western Blot (WB) — Western blot was performed using Anti-EphB2 Monoclonal Antibody (1A6C9) (Product # 37-1700) and 117, 110kDa bands corresponding to EphB2 was increased upon VEGF treatment in HUVEC. Also, the expression was highest in COLO 205 in comparison to SW480 and HT-29 as reported. Membrane enriched cell extracts (30 ug lysate) of HUVEC (Lane 1), HUVEC treated with VEGF (20ng/ml, 10 min) (Lane 2), COLO 205 (Lane 3), SW480 (Lane 4) and HT-29 (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

EphB2 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissues owing to their inherent genetic constitution. Relative expression of EphB2 was observed in COLO 205 in comparison to SW480 and HT-29 using EphB2 Monoclonal Antibody (1A6C9) (Product # 37-1700) in western blot. {RE}

EphB2 Antibody in Western Blot, Immunocytochemistry, Immunoprecipitation (WB, ICC/IF, IP) — Figure 6 EphB receptors Induce Tyrosine Phosphorylation of NrCAM at the FIGQY Motif, and Inhibit Ankyrin Recruitment in HEK293 cells. A. NrCAM was expressed with or without EphB2 or EphB2 kinase dead mutant (EphB2 KD) in transfected HEK293 cells, followed by immunoprecipitation with NrCAM antibodies and immunoblotting with phospho-FIGQY antibodies. EphB2 induced phosphorylation of NrCAM (200 kD) at FIGQY (p-FIGQY) compared to NrCAM alone or to co-expression of the EphB2 kinase dead mutant. Blots were stripped and reprobed with antibodies to NrCAM to indicate relative levels of NrCAM in the immunoprecipitates. B. NrCAM was expressed with or without EphB1, EphB2, or EphB3 in transfected HEK293 cells, followed by immunoprecipitation with NrCAM antibodies and immunoblotting with phospho-FIGQY antibodies. EphB1 (HA-tagged) or EphB2 (not HA-tagged) induced effective phosphorylation of NrCAM at FIGQY, whereas EphB3 (HA-tagged) had a weak effect. Blots were stripped and reprobed with antibodies to EphB2 or to the HA-epitope on EphB1 and EphB3. Each EphB receptor was co-immunoprecipitated efficiently with NrCAM. C. Lysates of the superior colliculus from WT, EphB1/3 double null, EphB1/2/3 triple null and constitutively active EphB2 (F620D) homozygous mice (P2-P3) were immunoprecipitated with antibodies to NrCAM, and immunoblotted with phospho-FIGQY or NrCAM antibodies. Bands were quantified using the threshold function of ImageJ. The numbers under the blots represent the normalized av

EphB2 Antibody in Western Blot, Immunoprecipitation (WB, IP) — Figure 1. Multiple guidance cue receptors interact with ribosomes. ( A ) Volcano plots showing statistically enriched proteins in DCC-IP and Nrp1-IP samples identified by permutation-based FDR-corrected t-test based on three biological replicates. The LFQ intensity of the DCC or Nrp1 pulldowns over IgG pulldowns are plotted against the -log10 p-value. FDR < 0.05; S0 = 2. ( B ) Gene enrichment analysis of statistically enriched proteins in the DCC and Nrp1 pulldown samples. The values in each circle denotes protein count. ( C-F ) Western blot validation of RP co-immunoprecipitation with DCC, Nrp1 and Robo2 but not with EphB2. Each Western blot was repeated 2 to 4 times, representative images are shown. ( G-J ) Relative 18S and 28S ribosomal RNA abundance after control (IgG) pulldown or receptors pulldowns shows enrichment of rRNA in DCC, Nrp1, and Robo2 but not EphB2 pulldowns (unpaired two-tailed t-test; three biological replicates). Bars indicate means, error bars indicate standard deviation; *p<0.05. Figure 1--figure supplement 1. Multiple guidance cue receptors interact with ribosomes in SH-SY5Y cells. ( A-C ) Western blot validation of RP co-immunoprecipitation with DCC, Nrp1 and Robo2 in SH-SY5Y cells. Western blots were repeated 2 to 4 times, Rps4X Western blots are from one experiment, representative examples are shown. ( D-E ) Relative 18S and 28S ribosomal RNA abundance after control (IgG) pulldowns or receptor pulldowns shows enrichment of rRNA in DCC and Nrp1 IPs i

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500

View 5 publications 5 publications

Immunocytochemistry (ICC/IF)

View 2 publications 2 publications

ELISA (ELISA)

Assay-dependent

Immunoprecipitation (IP)

Assay-dependent

View 3 publications 3 publications

Miscellaneous PubMed (Misc)

View 2 publications 2 publications

Product Specifications

Species Reactivity

Chicken, Human

Published species

Human, Mouse

Host/Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

1A6C9

Immunogen

A synthetic peptide derived from the C-terminal region of the recombinant EphB2 protein.

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_2533302

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. EphB4 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.

Bioinformatics

Protein Aliases: cEK5; chicken embryo kinase 5; Developmentally-regulated Eph-related tyrosine kinase; EFL6; EFNB3; EK5; ELK-related tyrosine kinase; embryo kinase 5 protein CEK5; EPH B2; EPH tyrosine kinase 3; EPH-like kinase 5; EphB2/CTF1; EphB2/CTF2; EPHB3; Ephrin B1; Ephrin B2; Ephrin B3; Ephrin B4; Ephrin type-B receptor 2; EphrinB1; EphrinB2; EphrinB3; EphrinB4; EPLG8; ETK2; HEK-6; HEK2; LERK8; MGC87492; protein-tyrosine kinase HEK5; Renal carcinoma antigen NY-REN-47; TYRO6; Tyrosine-protein kinase receptor EPH-3; Tyrosine-protein kinase TYRO5

Gene Aliases: CAPB; CEK5; DRT; EK5; EPHB2; EPHT3; EPTH3; ERK; HEK5; PCBC; TYRO5

UniProt ID: (Human) P29323, (Chicken) P28693

Entrez Gene ID: (Human) 2048, (Chicken) 396513

Molecular Function: transmembrane signal receptor