Home DNA Modifying Enzymes and Cloning Technologies Products Apyrase

Apyrase
NEBU cloned at NEB recombinant 30 65 Heat

Apyrase (recombinant, E. coli) is a highly active ATP-diphosphohydrolase that catalyzes the sequential hydrolysis of ATP to ADP and ADP to AMP releasing inorganic phosphate

  • Enzyme of highest purity, efficiency and quality 
  • Tested in updated range of more stringent quality controls
  • Supplied at 10-fold higher concentration in the optimized storage buffer
  • Supplied with new optimal reaction buffer
  • Provided with updated, detailed characterization
  • Discovered and characterized new function: the ability to convert 5’ triphosphorylated RNA to monophosphorylated form, which can be ligated or removed by 5’ exonuclease
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Catalog # Concentration Size List Price Your Price Quantity
M0398S 500 units/ml 10 units $82.00
M0398L 500 units/ml 50 units $333.00
Please enter a quantity for at least one size
  • Apyrase (recombinant, E. coli) is a highly active ATP-diphosphohydrolase that catalyses the sequential hydrolysis of ATP to ADP and ADP to AMP releasing inorganic phosphate. It is a recombinant version of one of several isoforms of apyrase. It can also hydrolyse 5´ tri- and diphosphate ribonucleosides and deoxyribonucleosides to their respective 5´ monophosphates (1). Apyrase can catalyse the conversion of 5´ triphosphorylated RNA to 5´ monophosphorylated RNA by sequential removal of γ and β phosphates (2).

    Product Source

    Isolated from a strain of E. coli that carries the coding sequence for potato S. tuberosum apyrase (4).
    This product is related to the following categories:
    Phosphatases Products,
    RNA Modification

    • Reagents Supplied

      The following reagents are supplied with this product:

      NEB # Component Name Component # Stored at (°C) Amount Concentration
      • M0398S     -20    
          Apyrase M0398SVIAL -20 1 x 0.02 ml 500 units/ml
          Apyrase Reaction Buffer B0398SVIAL -20 1 x 1.25 ml 10 X
      • M0398L     -20    
          Apyrase M0398LVIAL -20 1 x 0.1 ml 500 units/ml
          Apyrase Reaction Buffer B0398SVIAL -20 1 x 1.25 ml 10 X

    • Properties & Usage

      Unit Definition

      One unit is defined as the amount of enzyme that catalyses the release of 1 μmol of inorganic phosphate from ATP (1 mM, NEB #P0756) in 1X Apyrase Reaction Buffer in 1 minute at 30°C in a total reaction of 50 μl.

      Reaction Conditions

      1X Apyrase Reaction Buffer
      Incubate at 30°C

      1X Apyrase Reaction Buffer
      20 mM MES
      50 mM NaCl
      5 mM CaCl2
      1 mM DTT
      0.05% Tween® 20
      (pH 6.5 @ 25°C)

      Storage Buffer

      20 mM MES
      50 mM NaCl
      0.1 mM CaCl2
      1 mM DTT
      0.1% Tween® 20
      50% Glycerol
      pH 6.5 @ 25°C

      Heat Inactivation

      65°C for 20 minutes

      Molecular Weight

      Apparent: 47 kDa

      Specific Activity

      3,000 units/mg

    • Advantages and Features

      Features

      • Enzyme of highest purity, efficiency and quality 
      • Tested in updated range of more stringent quality controls
      • Supplied at 10-fold higher concentration in the optimized storage buffer
      • Supplied with new optimal reaction buffer
      • Provided with updated, detailed characterization
      • Discovered and characterized a new function: the ability to convert 5’ triphosphorylated RNA to monophosphorylated form, which can be ligated or removed by 5’ exonuclease

      Application Features

      • Highly efficient degradation of ATP to AMP.
      • Removal of deoxynucleotides in DNA pyrosequencing between cycles (3).
      • Conversion of 5´ triphosphorylated RNA to ligatable monophosphorylated form that can be used for 5´ RNA adaptor ligation.
      • Conversion of 5´ triphosphorylated RNA to 5´ exonuclease XRN-1 (NEB #M0338) sensitive monophosphorylated RNA.

    • Related Products

      Companion Products

    • Product Notes

      1. Apyrase has a higher ratio of activity for ATP:ADP (14:1).
      2. Apyrase is a calcium-activated enzyme. It is approximately 50% active when Mg2+ substitutes Ca2+ in Apyrase Reaction Buffer.
      3. As a metal-dependent enzyme, Apyrase can be inhibited by EGTA and EDTA.
      4. The activity of Apyrase is approximately 30% in NEBuffers r1.1, r2.1, r3.1 and rCutSmart™ Buffer.
      5. Apyrase does not remove 5´ caps from eukaryotic mRNA.

    • References

      1. Kettlun, A.M. et al. (2005). Phytochemistry. 66, 975-982.
      2. Schildkraut, I., Schildkraut, E. and Barshevsky, T., New England Biolabs, Inc. Unpublished observation
      3. Hornung, V. et al . (2006). Science. 314, 994-997.
      4. Schildkraut, I. and Schildkraut, E., New England Biolabs, Inc. Unpublished observation

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