Eph receptor B2 Recombinant Rabbit Monoclonal Antibody [PSH14-91]

Overview

Product Name

Eph receptor B2 Recombinant Rabbit Monoclonal Antibody [PSH14-91] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human Eph receptor B2 aa 1-543.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IP

Target Molecular Weight

Predicted band size: 117 kDa

Positive Control

COLO205 cell lysate, Saos-2 cell lysate, SW480 cell lysate, COLO205.

Conjugation

unconjugated

Clone Number

PSH14-91

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

WB
1:5,000

IF-Cell
1:100

IP
1-2μg/sample

Target

Function

Ephrin type-B receptor 2 is a protein that in humans is encoded by the EPHB2 gene. 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 two 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 is a receptor for ephrin-B family members.

Background References

1. Liu W et al. The Roles of EphB2 in Cancer. Front Cell Dev Biol. 2022 Feb

2. Xiao Y et al.Hepatocytes demarcated by EphB2 contribute to the progression of nonalcoholic steatohepatitis. Sci Transl Med. 2023 Feb

Subcellular Location

Cell membrane, Cell projection, axon, dendrite.

UNIPROT

SwissProt: P29323 Human

Synonyms

cek5 antibody

Developmentally regulated EPH related tyrosine kinase antibody

DRT antibody

EK5 antibody

ELK related protein tyrosine kinase antibody

Eph receptor B2 antibody

EPH tyrosine kinase 3 antibody

EPH-like kinase 5 antibody

EPHB2 antibody

EPHB2_HUMAN antibody

Expand

Images

☑ Relative expression (RE)

Western blot analysis of Eph receptor B2 on different lysates with Rabbit anti-Eph receptor B2 antibody (HA751535) at 1/5,000 dilution.

Lane 1: COLO205 cell lysate
Lane 2: K-562 cell lysate (negative)
Lane 3: Saos-2 cell lysate
Lane 4: SW480 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 117 kDa
Observed band size: 130 kDa

Exposure time: Lane 1-3: 20 seconds; Lane 4: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751535) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Relative expression (RE)

Immunocytochemistry analysis of COLO205 (positive) and K-562 (negative) labeling Eph receptor B2 with Rabbit anti-Eph receptor B2 antibody (HA751535) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Eph receptor B2 antibody (HA751535) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Eph receptor B2 was immunoprecipitated from 0.2 mg COLO205 cell lysate with HA751535 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751535 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: COLO205 cell lysate (input)
Lane 2: HA751535 IP in COLO205 cell lysate
Lane 3: Rabbit IgG instead of HA751535 in COLO205 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"