• My NEB
  • Print
  • PDF
  • T-Cell Protein Tyrosine Phosphatase (TC PTP)

    Discontinued Date

    01/01/2013
    recombinant unique buffer heat inactivation
    Catalog #SizeConcentrationPriceQtyAdd to Cart
      
    Categories:
    Discontinued Products

    Description



    T-Cell Protein Tyrosine Phosphatase (TC PTP) is a phospho tyrosine-specific protein phosphatase. It is a truncated form of the human T-Cell protein tyrosine phosphatase (residues 1-317) which lacks a C-terminal regulatory domain (1,2).

    Highlights

  • Protein phosphatase exclusively specific to phospho-tyrosine residues in proteins.
  • Human, Recombinant (E. coli)
  • Supplied with 10X Reaction Buffer
  • Product Source

    Isolated from a strain of E. coli that carries a clone expressing the T-Cell protein tyrosine phosphatase under the control of the T7 expression system (kindly provided by Dr. D. Bardford).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    NEBuffer for Protein Tyrosine Phosphatases (PTP)-2010X

    Advantages and Features

    Applications

    • TC PTP can be used to release phosphate groups specifically from phospho-tyrosine residues in proteins. Note that different proteins are dephosphorylated at different rates.

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme that hydrolyzes 1 nmol of p-Nitrophenyl Phosphate (50 mM) (NEB #P0757) in 1 minute at 30°C in a total reaction volume of 50 μl.

    Reaction Conditions

    1X NEBuffer for Protein Tyrosine Phosphatases (PTP)
    Incubate at 30°C

    1X NEBuffer for Protein Tyrosine Phosphatases (PTP):
    50 mM Tris-HCl
    100 mM NaCl
    2 mM Na2EDTA
    5 mM DTT
    0.01% Brij 35
    pH 7.5 @ 25°C

    Storage Temperature

    -70°C

    Storage Conditions

    50 mM HEPES
    100 mM NaCl
    5 mM DTT
    2 mM EDTA
    50% Glycerol
    0.01% Brij 35
    pH 7.0 @ 25°C

    Heat Inactivation

    65°C for 60 min

    Molecular Weight

    Theoretical: 37 kDa

    Specific Activity

    40,000 units/mg

    Storage Notes

    • Avoid repeated freeze/thaw cycles.
    • Can be stored for 2 weeks or less at -20°.

    Shipping Notes

    • Ships on dry ice

    Quality Control

    Quality Assurance Statement

    • TC PTP has been purified to >95% homogeneity as determined by SDS-PAGE and Coomassie Blue staining.

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Protease Activity (SDS-PAGE):

      The product is tested for protease activity by incubation with a standard mixture of proteins resulting in no detectable degradation of the proteins as determined by SDS-PAGE.

    • Protein Serine/Threonine Phosphatase (PSP) Activity (Radioactivity Release):
      The Phosphatase is tested in a reaction with myelin basic protein phosphorylated exclusively on serine/threonine residues in the presence of [33P]-ATP. After incubation no PSP activity is detected by release of radioactivity.

    Notes

    1. The following information can be used as suggested initial conditions for dephosphorylation of proteins with TC PTP.
    2. 1 unit of TC PTP removes ~100% of phosphates (0.5 nmol) in phosphorylated myelin basic protein (phospho-MyBP, 18.5 kDa) in 30 minutes in a 50 µl reaction. The concentration of phospho-MyBP is 10 µM with respect to phosphate.
    3. The Protein Tyrosine Phosphatase (PTP) activity of TC PTP is assessed on MyBP phosphorylated exclusively on tyrosine residues with Abl Protein Tyrosine Kinase.
    4. Optimal incubation times and enzyme concentrations must be determined empirically for each particular substrate.
    5. TC PTP is inhibited by vanadate (2).
    6. If the source of phosphorylated protein is a crude extract of cells or tissue, it is very important to include the appropriate protease inhibitors in the lysis buffer and to use shorter incubation time for dephosphorylation.
    7. The following levels of inhibition of TC PTP (1 unit) are found when the reaction buffer is supplemented with: 
      • 1 mM Sodium Orthovanadate (NEB #P0758) (3): 100% 
      • 50 mM Sodium Fluoride (NEB #P0759): no 
      • 1% Triton X-100: no 
      • 0.4% Nonidet P-40: no 
      • 0.025% Tween 20: no 
      • 0.5M NaCl : 10% 
      • ATP Mix (10 mM MgCl2, 0.1 mM ATP): no 
      • Protease inhibitor cocktail*: 10% 
      *Pepstatin A, leupeptin and aprotinin, 10 µg/ml each, 0.5 mM PMSF and 1 mM benzamidine.

    References

    1. Cool, D.E. et al. Proc. Natl. Acad. Sci. USA. 86, 5257-5261.
    2. Zander, N.F. et al. Biochemistry. 30, 6964-6970.
    3. Ruzzene, M. et al. Eur. J. Biochem.. 211, 289-295.
    4. Gordon, J.A. Methods Enzymol.. 201, 477-482.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

    Selection Tools