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 (Catalog # http://www.rndsystems.com/product_results.aspx?k=ES006">ES006) by detecting release of phosphate (Catalog # http://www.rndsystems.com/product_results.aspx?k=DY996">DY996).
Recombinant Human Active SHP-2 Protein, CF
R&D Systems | Catalog # 1894-SH
Key Product Details
- R&D Systems E. coli-derived Recombinant Human Active SHP-2 Protein (1894-SH)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
Accession Number
Applications
Product Specifications
Source
Thr2-Arg593, with an N-terminal 6-His tag
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >3 µmol/min/mg, measured under the described conditions.
Formulation, Preparation, and Storage
1894-SH
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: SHP-2
References
- Zhao, Z. et al. (1994) J. Biol. Chem. 269:8780.
- Clemmons, D.R. and Maile, L.A. (2005) Mol. Endocrinol. 19:1.
- von Wichert, G. et al. (2003) EMBO J. 22:5023.
- Maroun, C.R. et al. (2000) Mol. Cell. Biol. 20:8513.
- Sugimoto, S. et al. (1993) J. Biol. Chem. 269:22771.
Long Name
Alternate Names
Gene Symbol
UniProt
Additional SHP-2 Products
Product Documents for Recombinant Human Active SHP-2 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Active SHP-2 Protein, CF
For research use only
Citations for Recombinant Human Active SHP-2 Protein, CF
Powered by Bioz
Customer Reviews for Recombinant Human Active SHP-2 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Active SHP-2 Protein, CF and earn rewards!
Have you used Recombinant Human Active SHP-2 Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Protocols
View specific protocols for Recombinant Human Active SHP-2 Protein, CF (1894-SH):
- Assay Buffer: 25 mM HEPES, 0.02% Brij-35, 1 mM DTT, pH 7.5
- Recombinant Human SHP-2 (rhSHP-2) (Catalog # 1894-SH)
- Substrate: Asp-Ala-Asp-Glu-Tyr(PO3)-Leu-Ile-Pro-Gln-Gln-Gly (Catalog # ES006)
- Malachite Green Phosphate Detection Kit (Catalog # DY996)
- 96 well Clear Plate (Costar, Catalog # DY990)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Prepare a Phosphate Standard curve by adding 10 µL of the 1 M Phosphate Standard to 990 µL of Assay Buffer for 10 mM stock. Continue by adding 10 µL of the 10 mM phosphate stock to 990 µL of Assay Buffer for 100 µM stock. (this is the first dilution to use as a standard).
- Perform six additional one-half dilutions of the 100 µM Phosphate stock.
- Load 50 µL of each standard dilution to empty wells of a clear plate. Include a curve blank containing 50 µL Assay Buffer. The standard curve has a range of 0.078-5 nmol per well.
- Dilute rhSHP-2 to 0.2 µg/mL in Assay Buffer.
- Dilute Substrate to 400 μM in Assay Buffer.
- Load 25 µL of 0.2 µg/mL rhSHP-2 to empty wells. For controls, add 25 µL of Assay Buffer.
- Start reaction by adding 25 µL of 400 μM Substrate to enzyme and control wells.
- Cover plate with plate sealer and incubate at 37 °C for 30 minutes.
- After incubation, add 30 µL of Malachite Green reagent A to all wells used. Mix briefly.
- Add 100 μL of deionized water to all wells. Mix briefly.
- Add 30 µL of Malachite Green reagent B to all wells. Mix and incubate at room temperature for 20 minutes.
- Read plate at 620 nm in endpoint mode.
- Calculate specific activity:
|
Specific Activity (μmol/min/mg) = |
Phosphate released* (nmol) x (0.001 μmol/1 nmol) |
| Incubation time (min) x amount of enzyme (mg) |
*Derived from the phosphate standard curve using linear or 4-parameter fitting and adjusted for control.
Per Reaction:
- rhSHP-2: 0.000005 mg
- Substrate: 200 μM
FAQs for Recombinant Human Active SHP-2 Protein, CF
-
Q: Does Recombinant Human Active SHP-2 Protein, CF (Catalog # 1894-SH) require an activation step?
A: Based on in house testing, Catalog # 1894-SH does not require an activation step to meet our specific activity specifications of >3 µmol/min/mg. However, if a higher specific activity is required for a particular application, an activation step may be recommended. We suggest referring to the available literature on the product-specific page.
