Recombinant Human Active SHP-2 Protein, CF

Catalog # Availability Size / Price Qty
1894-SH-100
1894-SH-01M
Product Details
Citations (2)
FAQs
Supplemental Products
Reviews

Recombinant Human Active SHP-2 Protein, CF Summary

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to dephosphorylate a tyrosine residue in a peptide containing the EGFR Y992 phosphorylation site (Catalog # ES006). The specific activity is >3 µmol/min/mg, measured under the described conditions.
Source
E. coli-derived human SHP-2 protein
Thr2-Arg593, with an N-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
His
Predicted Molecular Mass
69 kDa
SDS-PAGE
63-70 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
  1. 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).
  2. Perform six additional one-half dilutions of the 100 µM Phosphate stock.
  3. 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.
  4. Dilute rhSHP-2 to 0.2 µg/mL in Assay Buffer. 
  5. Dilute Substrate to 400 μM in Assay Buffer.
  6. Load 25 µL of 0.2 µg/mL rhSHP-2 to empty wells. For controls, add 25 µL of Assay Buffer.
  7. Start reaction by adding 25 µL of 400 μM Substrate to enzyme and control wells.
  8. Cover plate with plate sealer and incubate at 37 °C for 30 minutes.
  9. After incubation, add 30 µL of Malachite Green reagent A to all wells used.  Mix briefly.
  10. Add 100 μL of deionized water to all wells.  Mix briefly.
  11. Add 30 µL of Malachite Green reagent B to all wells. Mix and incubate at room temperature for 20 minutes.
  12. Read plate at 620 nm in endpoint mode.
  13. 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
Reconstitution Calculator

Reconstitution Calculator

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.

=
÷

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 (Catalog # ES006) by detecting release of phosphate (Catalog # DY996).

References
  1. Zhao, Z. et al. (1994) J. Biol. Chem. 269:8780.
  2. Clemmons, D.R. and Maile, L.A. (2005) Mol. Endocrinol. 19:1.
  3. von Wichert, G. et al. (2003) EMBO J. 22:5023.
  4. Maroun, C.R. et al. (2000) Mol. Cell. Biol. 20:8513.
  5. Sugimoto, S. et al. (1993) J. Biol. Chem. 269:22771.
Long Name
Src Homology Region 2 Domain Phosphatase 2
Entrez Gene IDs
5781 (Human); 54390 (Mouse); 25622 (Rat)
Alternate Names
BPTP3; EC 3.1.3.48; MGC14433; protein tyrosine phosphatase, non-receptor type 11; protein tyrosine phosphatase-2; Protein-tyrosine phosphatase 1D; Protein-tyrosine phosphatase 2C; PTP1D; PTP-1D; PTP2C; PTP-2C; PTP2CNS1; PTPN11; SHP2; SHP-2; SHP2CFC; SHPTP2; SH-PTP2; SH-PTP2Noonan syndrome 1; SH-PTP3; tyrosine-protein phosphatase non-receptor type 11

Citations for Recombinant Human Active SHP-2 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
Filter your results:

Filter by:

  1. The tyrosine phosphatase SHP2 controls TGF?-induced STAT3 signaling to regulate fibroblast activation and fibrosis
    Authors: A Zehender, J Huang, AH Györfi, AE Matei, T Trinh-Minh, X Xu, YN Li, CW Chen, J Lin, C Dees, C Beyer, K Gelse, ZY Zhang, C Bergmann, A Ramming, W Birchmeier, O Distler, G Schett, JHW Distler
    Nat Commun, 2018;9(1):3259.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  2. Sepsis-induced cardiac mitochondrial dysfunction involves altered mitochondrial-localization of tyrosine kinase Src and tyrosine phosphatase SHP2.
    Authors: Zang Q, Martinez B, Yao X, Maass D, Ma L, Wolf S, Minei J
    PLoS ONE, 2012;7(8):e43424.
    Species: Rat
    Sample Types: Mitochondria
    Applications: Bioassay

FAQs

No product specific FAQs exist for this product, however you may

View all Proteins and Enzyme FAQs

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/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review