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| Download: | | MSDS PDF | |
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- Isolated from a recombinant source
- Supplied with 10X Reaction Buffer
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
Source:
This preparation is purified from K. lactis containing a clone of the potato apyrase gene (1).
Reagents Supplied:
Succinate Buffer for control purposes (10X)
Reaction & Storage Conditions
Unit Definition:
One unit is defined as the amount of enzyme that catalyzes the conversion of 1��mol of ATP to ADP in one minute at 30°C in a total reaction volume of 40 �l.
Unit Assay Conditions:
1X Succinate Buffer, 40�mM sodium succinate (pH 6.5), 4 mM CaCl2 and 1 mM ATP at 30°C.
Concentration:
50,000 milliunits/ml
Storage Conditions:
10 mM Tris-acetate
50 mM NaCl
0.1 mM CaCl2
0.1 mM DTT
50% Glycerol
0.1% Tween-20
Storage Temperature:
-20°C
Notes
General notes:
- 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.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Phosphatase Contamination:
After incubation of 1 unit of Apyrase with 0.05 �mol p-nitrophenol phosphate for 20 hours at 37°C, no phosphatase activity could be detected by spectrophotometric analysis.
Nuclease Contamination:
Incubation of 1 unit of Apyrase for 6 hours in the recommended assay buffer with Lambda-HindIII Digest and 100 bp DNA Ladder revealed no detectable endonuclease activity as determined by agarose gel electrophoresis.
Exonuclease Activity:
Incubation of 50 �l reaction containing 1 unit of Apyrase with 1 �g of a mixture of single and double-stranded [3H] E.�coli DNA (200,000�cpm/�g) for 6�hours at 37°C released <�0.5% of the total radioactivity.
References
- Ganatra, M., Hough, D. and Taron, C.H., unpublished observations.