Nicholas T Santoro

US Patent:

20030049705, Mar 13, 2003

Filed:

Sep 13, 2001

Appl. No.:

09/682517

Inventors:

Heidi Dodson - Novi MI, US
James Marks - Ann Arbor MI, US
Thomas McQuade - Ypsilanti MI, US
Maxine Santoro - Dexter MI, US
Nicholas Santoro - Dexter MI, US

International Classification:

C12Q001/48
C12Q001/42

US Classification:

435/015000, 435/021000

Abstract:

A method for measuring the activity of dual substrate enzymes and for identifying compounds that modulate the activity of dual substrate enzymes comprising first substrate, a macromolecule that is enzymatically modified with a radioactively-labeled portion of the second substrate, said method comprising: a. adding a capture resin to a buffered mixture of an enzyme, allowing the enzyme to catalyze transfer of the radiolabeled portion of the radiolabeled second substrate to the non-radiolabeled first substrate, radiolabeled first substrate, and a radiolabeled second substrate in the presence or absence of a test compound; b. removing unreacted radiolabeled second substrate; c. adding a scintillant resin to the enzyme-radiolabeled first substrate mixture; and d. measuring the amount of radiolabeled first substrate by scintillation counting, measuring the amount of radiolabeled first substrate by scintillation counting, and comparing the two measurements.