Used as an enzyme label for ELISA (2).
Unit Definition: One unit will liberate 1 nmol of inorganic orthophosphate per 10 min at pH 7.5 at 65°C.
Storage Conditions: store at -20°C.
Storage Buffer: 20 mM Tris-HCl (pH 7.5 at 22°C), 50 mM NaCl, 1 mM dithiothreitol, 1 mM EDTA and 50% (v/v) glycerol.
Assay Conditions: 40 mM Tris-HCl (pH 7.5 at 22°C), 40 mM potassium acetate, 40 mM imidazole, 1 mM sodium pyrophosphate and 2.5 mM MgCl2. Incubation is at 65°C for 10 min in a reaction volume of 100 µl.
Quality Control: All preparations are assayed for contaminating endonuclease, exonuclease and nonspecific RNase and single- and double-stranded DNase activities.
References:
1. Baykov, A. A., Kasho, V.N. and Avaeva, S.M. (1988) Anal. Biochem. 171, 271-276.2. Jerhoeven, J.A. (1986) J. Bacteriol. 168, 318-321.
3. Richter, O. M. and Schafer, G. (1992) Eur. J. Biochem. 209, 343-349.