Method and reagent kit for determining activity of...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C435S007100, C435S007200

Reexamination Certificate

active

06861225

ABSTRACT:
The invention relates to a method for determining activity of 5′-nucleotidase, in which a biological sample is incubated in a manner and under conditions known per se with a nucleotide pentose monophosphate as substrate, the liberated inorganic phosphate is converted into a colored complex by treating it with ammonium molybdate and a reducing agent, color intensity is measured by a known method, and from the measured value the activity of 5′-nucle-otidase or the amount of inorganic phosphate liberated within unit time, as a figure proportional to the activity of 5′-nucleotidase, is calculated with a known calculation and/or with the aid of a calibration curve. According to the invention 5′-AMP, 5′-CMP, 5′-UMP, 5′-GMP, 5′-IMP and 5′-TMP are used as nucleotide pentose monophosphates, and measurement is performed on all of these six substrates. The invention also relates to a reagent kit for performing the above method.

REFERENCES:
Paglia et al. The Journal of Biological Chemistry, vol. 250, No. 20, pp. 7973-7979, 1975.*
El-Aaser, Abdel-basset Anwer, et al., “Simultaneous Determination of 5′-Nucleotidase and Alkaline Phosphatase Activities in Serum,”Z. Klin. Chem. Klin. Biochem., vol. 13, No. 10, pp. 453-459 (1975).
Parthasarathi, K., et al., “Effect of Spike Disease on the 5′- and 3′-Nucleotidase Activities in Sandal Plants,”Experientia, vol. 37, No. 5, pp. 448-449 (1981).
Wood, Raymond John, et al., “Colorimetric Determination of Serum 5′-Nucleotidase without Deproteinization,”Clin. Chem., vol. 27, No. 3, pp. 464-465 (1981).
Fioretti, E., et al., “Spectrophotometric Assays for 5′-Nucleotidase, Using IMP, GMP and CMP as Substrates,”The Italian Journal of Biochemistry, vol. 21, No. 3, pp. 103-112 (May-Jun. 1972).
Bodansky, Oscar, et al., “5′-Nucleotidase,”Advances in Clinical Chemistry, vol 11, pp. 277-328 (1968).

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