METHOD FOR DETERMINING OF ACTIVITY OF HUMAN APURIN/APIRIMIDIN-ENDONUCLEASE Russian patent published in 2010 - IPC G01N33/573 C12Q1/68 

FIELD: medicine.

SUBSTANCE: to implement the method for determining of activity of human apurin/apirimidin-endonuclease the reaction is precipitated by mixing solutions of substrate and enzyme in ratio 1:1 and register changes in the fluorescence intensity of the substrate in time interval of 1-2000 sec. For these purposes in basin of fluorimetre substrate solutions are mixed (1.0×10^-6 M), Tris-HCl buffer and sample of AP-endonuclease of a human (1.0×10^-7 M, 1.0×10^-8 M, 1.0×10^-9 M, 1.0×10^-10 M). A double-stranded complex of complementary oligodeoxyribonucleotides in length of 13 or more units containing in a central position and the rest of 2-hydroxymethyl-W-hydroxy-tetrahydrofuran and bearing a dye fluorescein and dabtsil are used as substrates. Changes in fluorescence intensity of the substrate in the reaction mixture is registered in a spectral range of 500-550 nm, fluorescence excitation is performed on wavelength of 494 nm. Kinetic curves are processed by nonlinear regression method using the program DynaFit.

EFFECT: increasing of accuracy and reliability of determining activity of a compound, ability to renounce the use of radioactively labeled substrate and standardise the units of activity in accordance with the international system.

1 dwg, 1 ex