Human/Mouse Phospho-SHP‑2 (Y542) Antibody

Name: Human/Mouse Phospho-SHP‑2 (Y542) Antibody
Brand: R&D Systems
SKU: AF3790
Price: 159 USD
Availability: InStock

R&D Systems | Catalog # AF3790

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Citations (2)

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Rat

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Western Blot, Simple Western, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

View all SHP-2 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing human SHP-2 Y542 site

Specificity

Detects human and mouse SHP-2 when phosphorylated at Y542.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse Phospho-SHP‑2 (Y542) Antibody

Detection of Mouse Phospho-SHP‑2 (Y542) by Western Blot.

Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50 ng/mL Human PDGF-BB (Catalog # 220-BB) for 20 minutes. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse Phospho-SHP-2 (Y542) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3790), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-SHP-2 (Y542) at approximately 72 kDa (as indicated). The lysates were also probed for total SHP-2 with Human/Mouse/Rat SHP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1894). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse Phospho-SHP‑2 (Y542) by Simple Western^TM.

Simple Western lane view shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50 ng/mL Recombinant Human PDGF-BB (Catalog # 220-BB) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-SHP-2 (Y542) at approximately 72 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human/Mouse Phospho-SHP-2 (Y542) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3790). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.*Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

SHP‑2 in THP‑1 Human Cell Line.

SHP‑2 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line treated with PMA(positive staining) and THP‑1 human acute monocytic leukemia cell line (untreated; negative staining) using Rabbit Anti-Human/Mouse Phospho-SHP‑2 (Y542) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3790) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Human/Mouse Phospho-SHP‑2 (Y542) Antibody

Application

Recommended Usage

Immunocytochemistry

3-15 µg/mL
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line treated with PMA

Simple Western

10 µg/mL
Sample: NIH‑3T3 mouse embryonic fibroblast cell line treated with Recombinant Human PDGF‑BB (Catalog # 220-BB)

Western Blot

1 µg/mL
Sample: NIH‑3T3 mouse embryonic fibroblast cell line treated with Human PDGF (Catalog # 120-HD)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: SHP-2

Src-Homology domain-2 containing protein tyrosine Phosphatase 2 (SHP-2), also called protein tyrosine phosphatase, non-receptor type 11 (PTPN11), PTP1D, PTP2C, and SYP, is an enzyme that dephosphorylates tyrosine residues in proteins. The protein contains two Src homology 2 (SH2) domains, which both regulate the activity of the enzyme (1) and allow it to selectively bind to SH2 sites on proteins such as Dok1, IRS1, and the insulin receptor (2). SHP-2 plays a unique stimulatory role in cell signaling. Cells lacking SHP-2 have poor mobility because the hyper-phosphorylation of FAK and other proteins in the focal adhesion complex (3) prevents turnover of cellular attachment points. Without SHP-2, sustained ERK stimulation does not take place (4). The Y992 phosphorylation site of EGFR is a particularly good substrate for SHP-2 (5) and a phosphopeptide containing this sequence can be used to measure the activity of the enzyme (R&D Systems, Catalog # ES006) by detecting release of phosphate (R&D Systems, Catalog # DY996).