REAGENT FOR DETERMINATION OF ADENOSINE 5'-TRIPHOSPHATE Russian patent published in 2006 - IPC C12Q1/66 

FIELD: biochemistry, analytical biochemistry.

SUBSTANCE: reagent for quantitative determination of adenosine 5'-triphosphate (ATP) by bioluminescent method comprises luciferase isolated from recombinant E. coli cells comprising plasmid pLR with native firefly luciferase gene, or luciferase isolated from recombinant E. coli cells comprising plasmid with m-pLR with the Luciola migrelica mutant luciferase gene wherein histidine residue at the position 433 is replaced for tyrosine residue. Also, the reagent comprises magnesium sulfate, mixture of tris-(hydroxymethyl)-aminomethane and acetic acid as a buffer, mixture of ethylenediaminetetraacetate sodium, dithiothreitol, bovine serum albumin and trehalose as stabilizing agents and water. In the assay method of adenosine 5'-triphosphate the reagent provides enhancing sensitivity and reproducibility in determination of ATP and reducing consumption of enzyme due to increase of specific activity of the reagent. The reagent provides carrying out determination of adenosine 5'-triphosphate by bioluminescence intensity at 560-570 nm in using firefly Luceola migrelica native luciferase, and by bioluminescence intensity at 606-610 nm in using luciferase wherein histidine residue at the position 433 is replaced for tyrosine residue.

EFFECT: improved assay method, valuable properties of reagent.

1 dwg, 1 tbl, 4 ex