[EN] DETERMINATION OF NUCLEASE ACTIVITY<br/>[FR] DETERMINATION DE L'ACTIVITE DE NUCLEASE
申请人:HARBRON STUART
公开号:WO2000049172A1
公开(公告)日:2000-08-24
A method for detecting a nuclease enzyme is disclosed which comprises a method for detecting a nuclease enzyme comprising the steps: a) contacting said enzyme with a compound of formula RpX, wherein R is a 3' nucleosidyl derivative, p is a phospho radical, and X is an esterifiable moiety or, only if R is a 3' nicotinamide derivative, X is an esterifiable moiety or H, whereby ROH and pX are produced, and b) detecting said pX moiety or, only if R is a 3' nicotinamide derivative, detecting the pX moiety or the ROH moiety. In preferred embodiments the invention provides a method for detecting a nuclease enzyme that is free in solution, immobilised on a surface, or attached to a member of a specific binding pair. The method of the invention may thus be applied as a detection step in nucleic acid hybridisation assays, enzyme immunoassays and ligand:receptor binding assays. The invention provides a variety of methods for detecting the detectable moieties produced. These include fluorometric, colorimetric, and luminometric endpoints. Enzyme cycling and apoenzyme reactivation assays are also provided.