Recombinant Human PTEN Protein, CF

R&D Systems | Catalog # 847-PN

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant Human PTEN Protein (847-PN)
  • 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

E. coli

Accession Number

P60484

Applications

Inhibition Activity
View all PTEN Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human PTEN protein
Thr2-Val403, with a C-terminal 6-His tag

Purity

>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Thr2

Predicted Molecular Mass

48 kDa

SDS-PAGE

59 kDa, reducing conditions

Activity

Measured by its ability to dephosphorylate either diC8PIP3 or diC16PIP3.
The specific activity is >480 nmol/min/mg, as measured under the described conditions.

Formulation, Preparation, and Storage

847-PN
Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl, EDTA, Glycerol and Betamercaptoethanol.
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.

Background: PTEN

The tumor suppressor gene PTEN (phosphatase and tensin homolog deleted on chromosome 10), also known as MMAC1 (mutated in multiple advanced cancers 1), encodes a phosphatase that contains the catalytic signature motif (HCXXGXXRS/T) found in all members of the protein tyrosine phosphatase family. In vitro, the recombinant PTEN has both lipid phosphatase and protein phosphatase activities (1, 2). Interestingly, accumulating evidence has shown that the tumor suppressor activity of PTEN relies on its ability to dephosphorylate phosphatidylinositol (3, 4, 5)-triphosphate specifically at position 3 of the inositol ring (3). This activity reduces the levels of phosphatidylinositol (3, 4, 5)-triphosphate which is specifically produced from phosphatidylinositol (4, 5)-diphosphate by PI 3-kinase upon activation by a variety of stimuli. Therefore, PTEN antagonizes PI 3-kinase-induced downstream signaling events and cellular processes including cell growth, apoptosis and cell motility. In vivo, the importance of PTEN catalytic activity in its tumor suppressor functions is underscored by the fact that the majority of PTEN missense mutations detected in tumor specimens target the phosphatase domain and cause a loss in PTEN phosphatase activity (4).

References

  1. Maehama, T. and J. Dixon (1998) J. Biol. Chem. 273:13375.
  2. Das, S. et al. (2003) Proc. Natl. Acad. Sci. USA 100:7491.
  3. Myers, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:13513.
  4. Waite, K. and C. Eng (2002) Am. J. Hum. Genet. 70:829.

Long Name

Phosphatase and Tensin Homolog Deleted on Chromosome 10

Alternate Names

MMAC1

Entrez Gene IDs

5728 (Human); 19211 (Mouse); 50557 (Rat)

Gene Symbol

PTEN

UniProt

P60484 (Human)

Additional PTEN Products

Product Documents for Recombinant Human PTEN 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.

Download SDS

Product Specific Notices for Recombinant Human PTEN Protein, CF

This product is covered by one or more of the following U.S. patents: 6,262,242, 6,482,795, and U.S. patent application serial number 10/299,003.

For research use only

Citations for Recombinant Human PTEN Protein, CF

Customer Reviews for Recombinant Human PTEN Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human PTEN Protein, CF and earn rewards!

Have you used Recombinant Human PTEN 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
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human PTEN Protein, CF (847-PN):

Materials
  • Assay Buffer: 100 mM Tris, 2 mM DTT, 0.05% (v/v) NP-40 Substitute (Sigma, Catalog # 74385), pH 8.5
  • Vesicle Buffer: 10 mM HEPES, 1 mM EGTA and 1 mg/mL BSA, pH 7.5
  • Recombinant Human PTEN (rhPTEN) (Catalog # 847-PN)
  • Substrate: diC16PIP3 (Echelon, Catalog # P-3916)
  • Lipids: dioleoyl phosphatidylcholine (DOPC), and dioleoyl phosphatidylserine (DOPS) (Avanti Polar Lipids, Catalog # 790595)
  • Malachite Green Phosphate Detection Kit (R&D Systems, Catalog # DY996)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Prepare a solution of 12.5 mg/mL DOPC, 12.5 mg/mL DOPS in Vesicle Buffer.
  2. Form Phospholipid Vesicles by combining substrate and lipids in Vesicle Buffer to final concentrations of 0.3 mM diC16PIP3, 625 μg/mL DOPC, 625 μg/mL DOPS. Mix vigorously.
  3. Dilute rhPTEN to 0.25 ng/μL in Assay Buffer.
  4. Dilute Phospholipid Vesicles to 0.09 mM diC16PIP3, 187.5 μg/mL DOPC, 187.5 μg/mL DOPS in Assay Buffer.
  5. Prepare a standard curve from the 1 M Phosphate Standard supplied in the malachite green phosphate detection kit. First, add 10 µL of the 1 M Phosphate Standard to 990 µL of Assay Buffer for a 10 mM stock. Then, add 10 µL of the 10 mM phosphate stock to 990 µL of Assay Buffer for a 100 µM stock. (This is the first dilution to use as a standard.) Finally, perform six additional two-fold serial dilutions of the 100 µM phosphate stock.
  6. Load 100 μL of each point of the curve to wells in triplicate.
  7. To the experimental wells, add 80 µL of 0.25 ng/μL rhPTEN. To the control wells, add 80 µL of Assay Buffer.
  8. Start the reaction by adding 20 µL of dilute Phospholipid Vesicles.
  9. Incubate at 37 ºC for 15 minutes.
  10. Add 20 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
  11. Add 20 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  12. Read plate at 620 nm (absorbance) in endpoint mode.
  13. Calculate specific activity:

     Specific Activity (nmol/min/mg) =

 Phosphate released* (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 wells.

Per Reaction:
  • rhPTEN: 20 ng = 0.00002 mg
  • Substrate: 18 µM
  • Standard Curve: 0.156, 0.313, 0.625, 1.25, 2.5, 5 and 10.0 nmol

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes