Recombinant Human TC-PTP (aa 2-314) Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human TC-PTP (aa 2-314) Protein, CF Summary

Details of Functionality
Measured by its ability to dephosphorylate a tyrosine residue in a peptide containing the EGFR Y992 phosphorylation site (Catalog # ES006). The specific activity is >30 μmol/min/mg, as measured under the described conditions.
Source
E. coli-derived human TC-PTP protein
Thr2-Asn314, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
PTPN2
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
37 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
38 kDa, reducing conditions
Publications
Read Publications using
1930-PT in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES, NaCl, Glycerol and Betamercaptoethanol.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 10 mM HEPES, 0.1 mM EDTA, 0.1 mM EGTA, 0.5 mg/mL BSA, 1 mM dithothreitol, pH 7.5
  • Recombinant Human TC‑PTP (rhTC-PTP) (Catalog # 1930-PT)
  • 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 # 92592) or equivalent
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  • Phosphate Standard: 1 M KH2PO4
  1. Dilute rhTC-PTP to 0.025 µg/mL in Assay Buffer.
  2. Load 40 µL of diluted rhTC-PTP to a plate. Include Substrate Blanks containing 40 µL Assay Buffer in place of rhTC-PTP.
  3. Dilute the Substrate in Assay Buffer to 1 mM and add 10 µL to the wells.
  4. Cover the plate with parafilm or a plate sealer and incubate at 30 °C for 30 minutes.
  5. Prepare a standard curve from the 1M Phosphate Standard by adding 10 µL of the 1 M Phosphate Standard to 990 µL of assay buffer for a 10 mM stock. Continue by adding 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.) Perform six additional one-half serial dilutions of the 100 µM Phosphate stock. The standard curve has a range of 0.078 to 5 nmol per well.
  6. Load 50 µL of the standards and blanks to the plate.
  7. Add 10 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
  8. Add 10 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  9. Read plate at 620 nm (absorbance) in endpoint mode.

10.  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 Substrate Blank.

Per Well:
  • rhTC-PTP: 0.000001 mg (1 ng)
  • Substrate: 200 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TC-PTP (aa 2-314) Protein, CF

  • protein tyrosine phosphatase, non-receptor type 2
  • PTP2
  • PTPN2
  • PTPTPTN2
  • T-cell protein tyrosine phosphatase
  • T-cell protein-tyrosine phosphatase
  • TCELLPTP
  • TCPTP
  • TC-PTP
  • TCPTPEC 3.1.3.48
  • tyrosine-protein phosphatase non-receptor type 2

Background

T-cell protein tyrosine phosphatase (TC-PTP), also known as PTPT and PTPN2, is an enzyme that removes phosphate groups covalently attached to tyrosine residues in proteins. This enzyme has two C-terminal end splice variants with distinctly different subcellular localizations. The shorter 45 kilodalton isoform is exclusively nuclear in resting cells, but redistrubutes to the cytosol upon stimulation with growth factors (1) and cellular stress (2). The longer 48 kilodalton isoform is exclusively found in the endoplasmic reticulum (3) and seems to have distinctly different physiologic substrates from the smaller isoform (1, 4). Although found in many cell types and tissues, TC-PTP is particularly prominent in hemopoietic cell types (5, 6). Knockout mice lacking TC-PTP are born viable but die 3 to 5 weeks after birth of erythropoietic and lymphopoietic deficits (7), indicating a critical role for TC-PTP in bone marrow maturation. TC-PTP will dephosphorylate a wide range of phosphoproteins, such as p52Shc (6) and receptors for EGF (1), Insulin (8) and growth hormone (6). The recombinant protein lacks the C-terminal 100 amino acids that determine intracellular localization but is fully active (9).

  1. Tiganis, T. et al. (1999) J. Biol. Chem. 274:27768.
  2. Lam, M.H. et al. (2001) J. Biol. Chem. 276:37700.
  3. Lorenzen, J.A. et al. (1995) J. Cell Biol. 131:631.
  4. Tiganis, T. et al. (1998) Mol. Cell. Biol. 18:1622.
  5. Cool, D.E. et al. (1989) Proc. Natl. Acad. Sci. USA 86:5257.
  6. Pasquali, C. et al. (2003) Mol. Endocrinol. 17:2228.
  7. You-Ten, K.E. et al. (1997) J. Exp. Med. 186:683.
  8. Galic, S. et al. (2003) Mol. Cell. Biol. 23:2096.
  9. Cool, D.E. et al. (1990) Proc. Natl. Acad. Sci. USA 87:7280.

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1930-PT
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Publications for TC-PTP/PTPN2 (1930-PT)(3)

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Bioinformatics

Gene Symbol PTPN2
Uniprot