Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Tuesday March 02, 2021

Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP508(2)Y] to SHP2 (phospho Y542)
Suitable for: ICC/IF, WB, IP
Reacts with: Mouse, Human

Product name

Anti-SHP2 (phospho Y542) antibody [EP508(2)Y]
See all SHP2 primary antibodies
Description

Rabbit monoclonal [EP508(2)Y] to SHP2 (phospho Y542)
Host species

Rabbit
Specificity

ab62322 detects SHP2 phosphorylated at Tyrosine 542.

Tested Applications & Species

Application	Species
ICC/IF	Human
IP	Mouse
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- Treated NIH/3T3 cell lysate.
General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP508(2)Y
Isotype

IgG
Research areas

Immunoprecipitation - Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322)

ab62322 (purified) at 1:30 dilution (2µg) immunoprecipitating SHP2 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) treated with 50ng/ml PDGF for 40min whole cell lysate 10µg
Lane 2 (+): ab62322 & NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab62322 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
Western blot - Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322)

All lanes : Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322) at 1/1000 dilution (Purified)

Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 40ng/ml PDGF for 40 minutes whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 40ng/ml PDGF for 40 minutes then incubated with phosphatase

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa
Immunocytochemistry/ Immunofluorescence - Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322)

Ab62322 staining SHP2 in THP-1 cells ( Human monocytic leukaemia cell line) by ICC/IF (immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with purified ab62322 at 8.8 μg/ml. Secondary antibody used was AlexaFluor^®488 Goat anti-Rabbit (ab150077) at 2 μg/ml. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)( ab195889) used as counterstain at 2.5 μg/ml . DAPI was used as nuclear counterstain. Confocal image showing the expression was increased after treatment with TPA 200nM for 24h and PDGF 50ng/ml for 30min, the signal decreased after treatment with Lambda Protein Phosphatase 31℃ for 2h.

This image was generated using the unpurified version of the product.
Western blot - Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322)

All lanes : Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322) at 1/200 dilution

Lane 1 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates
Lane 2 : THP-1 (Human monocytic leukemia monocyte) treated with Phorbol-12-myristate-13-acetate and platelet-derived growth factor.
Lane 3 : THP-1 (Human monocytic leukemia monocyte) treated with Phorbol-12-myristate-13-acetate and platelet-derived growth factor.Then the membrane was incubated with phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Blocking and diluting buffer used was 5% NFDM/TBST.

Purified batch of ab62322 was used.

This image was generated using the unpurified version of the product.
Western blot - Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322)

All lanes : Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322) at 1/50000 dilution

Lane 1 : NIH/3T3 cell lysates; untreated
Lane 2 : NIH/3T3 cell lysates; treated with PDGF

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : goat anti-rabbit HRP conjugated, at 1/2000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Beta Tubulin has been included as a loading control.

This image was generated using the unpurified version of the product.
Anti-SHP2 (phospho Y542) antibody [EP508(2)Y] (ab62322)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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