Apyrase |NEB酶试剂 New England Biolabs

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产品信息

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).

产品来源

Isolated from a strain of E. coli that carries the coding sequence for potato S. tuberosum apyrase (4).

产品类别:
Phosphatases Products,
RNA Modification

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • 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

  • 特性和用法

    单位定义

    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.

    反应条件

    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)

    贮存溶液

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

    热失活

    65°C for 20 minutes

    分子量

    实际: 47 kDa

    比活度

    3,000 units/mg

  • 优势和特性

    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

    应用特性

    • 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.

  • 相关产品

    相关产品

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    • Monarch® PCR & DNA 纯化试剂盒（5 μg）

  • 注意事项

    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.

  • 参考文献

    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

FAQs & 问题解决指南

  • FAQs

    1. What is the activity of Apyrase in other NEBuffers?
    2. Can Mg2+ be substituted for Ca2+ in Apyrase reaction buffer?