Eph receptor A2 Recombinant Rabbit Monoclonal Antibody [PSH09-21]

Overview

Product Name

Eph receptor A2 Recombinant Rabbit Monoclonal Antibody [PSH09-21] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human EPHA2 aa 10-550 / 976

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IP, FC

Target Molecular Weight

Predicted band size: 108 kDa

Positive Control

A549 cell lysate, HEK-293 cell lysate, HeLa cell lysate, A549, HeLa.

Conjugation

unconjugated

Clone Number

PSH09-21

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
1:2,000

IF-Cell
1:100

IP
1-2μg/sample

FC
1:1,000

Target

Function

This gene belongs to the ephrin receptor subfamily of the protein-tyrosine kinase family. EPH and EPH-related receptors have been implicated in mediating developmental events, particularly in the nervous system. Receptors in the EPH subfamily typically have a single kinase domain and an extracellular region containing a Cys-rich domain and 2 fibronectin type III repeats. The ephrin 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. This gene encodes a protein that binds ephrin-A ligands.

Background References

1. Xiaojun S ,Ryan L ,J C H , et al.Time-resolved live-cell spectroscopy reveals EphA2 multimeric assembly.[J].Science (New York, N.Y.),2023,eadg5314-eadg5314.

2. Qiu P ,Ling C ,Wei W , et al.EPH receptor A2 governs a feedback loop that activates Wnt/β-catenin signaling in gastric cancer.[J].Cell death & disease,2018,9(12):1146.

Subcellular Location

Cell membrane, Cell projection, ruffle membrane, lamellipodium membrane, Cell junction, focal adhesion.

UNIPROT

SwissProt: P29317 Human

SwissProt: Q03145 Mouse

Entrez Gene: 366492 Rat

Synonyms

ARCC2 antibody

AW545284 antibody

CTPA antibody

CTPP1 antibody

CTRCT6 antibody

EC 2.7.10.1 antibody

Eck antibody

Eph receptor A2 antibody

EPHA2 antibody

EPHA2_HUMAN antibody

Expand

Images

Western blot analysis of Eph receptor A2 on different lysates with Rabbit anti-Eph receptor A2 antibody (HA751275) at 1/2,000 dilution.

Lane 1: A549 cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: HeLa cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 108 kDa
Observed band size: 130 kDa

Exposure time: 59 seconds; 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 (HA751275) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of A549 cells labeling Eph receptor A2 with Rabbit anti-Eph receptor A2 antibody (HA751275) 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 A2 antibody (HA751275) 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 A2 was immunoprecipitated from 0.2 mg A549 cell lysate with HA751275 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751275 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: A549 cell lysate (input)
Lane 2: HA751275 IP in A549 cell lysate
Lane 3: Rabbit IgG instead of HA751275 in A549 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 59 seconds; ECL: K1801
Flow cytometric analysis of A549 cells labeling Eph receptor A2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751275, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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