• My NEB
  • Print
  • PDF
  • Apyrase

    Description

    Apyrase is an ATP diphosphohydrolase. It catalyses the removal of the gamma phosphate from ATP and the beta phosphate from ADP. The phosphate from AMP is not removed.

    10 units = 10,000 milliunits

    Highlights

  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
  • Product Source

    This preparation is purified from K. lactis containing a clone of the potato apyrase gene (1).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    10X Succinate Buffer (for control purposes)10X

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme that catalyzes the conversion of 1 µmol of ATP to ADP in one minute at 30°C in a total reaction volume of 50 µl.

    Reaction Conditions

    1X Succinate Buffer (for control purposes)

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-acetate
    50 mM NaCl
    0.1 mM CaCl2
    0.1 mM DTT
    50% Glycerol
    0.1% Tween® 20
    pH 6.5 @ 25°C

    Unit Assay Conditions

    1X Succinate Buffer, 40 mM sodium succinate (pH 6.5), 4 mM CaCl2 and 1 mM ATP at 30°C.

    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.
    • 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.

    Notes

    1. The activity of Apyrase at pH 7.5 in a Tris Buffer is approximately 80% of the activity at pH 6.5. Magnesium can substitute for calcium in the reaction. The ratio of ATPase:ADPase is 12:1 with this preparation of Apyrase.

    References

    1. Ganatra, M., Hough, D. and Taron, C.H. Unpublished observation

    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.