Product Class: Other

Apyrase
NEBU cloned at NEB recombinant 30 65 Heat

Product Introduction

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
Catalog # Size Concentration
M0398S 10.0 units 500 units/ml
M0398L 50.0 units 500 units/ml

Product Information

Description

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

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

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.

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

FAQs & Troubleshooting

FAQs

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

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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