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  • Thermostable Inorganic Pyrophosphatase

    Description

    Inorganic pyrophosphatase (PPase) catalyzes the hydrolysis of inorganic pyrophosphate to form orthophosphate:
    P2O7-4 + H2O -> 2HPO4-2.

    Highlights

    • Retains 100% activity after incubation at 100°C for 4 hours

    Product Source

    An E. coli strain carrying a plasmid encoding a pyrophosphatase from the extreme thermophile Thermococcus litoralis.

    Properties and Usage

    Unit Definition

    One unit is the amount of enzyme that will generate 1 μmol of phosphate per minute from inorganic pyrophosphate under standard reaction conditions (a 10 minute reaction at 75°C in 50 mM Tricine [pH 8.5] , 1 mM MgCl2, 0.32 mM PPi, reaction volume of 0.5 ml).

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM Tris-HCl
    100 mM KCl
    1 mM DTT
    0.1 mM EDTA
    50% Glycerol
    pH 8.0 @ 25°C

    Heat Inactivation

    No

    Quality Control

    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.
    • dNTPase Activity:
      The Pyrophosphatase is tested in a reaction containing a mix of dNTP's. After incubation the amount of inorganic phosphatase released is determined by comparison to a standard curve of known phosphate concentrations.
    • Endonuclease Activity (Nicking):
      The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
    • Exonuclease Activity (Radioactivity Release):
      The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
    • Non-Specific DNase Activity (16 hour):
      The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
    • Phosphatase Activity (PNPP):
      The product is tested in a reaction containing a p-nitrophenyl phosphate (PNPP), a chromogenic substrate for most phosphatases. After incubation there is no detectable phosphatase activity as determined by spectrophotometric analysis of released p-nitrophenol at 405 nm.

    References

    1. Heinonen, J.K. and Lahti, R.J. (1981). Analytical Biochemistry. 113, 313-317.

    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.
    1. What is the activity of TIPP at 37°C?