Rapid assay for radioactive determination of protein kinase acti

Details Rapid assay for radioactive determination of protein kinase acti Rapid assay for radioactive determination of protein kinase acti

1994-04-08

1996-07-23

Wityshyn, Michael G.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving transferase

435 4, 435 21, 435194, 436504, 436804, C12Q 148, A61K 5100

Patent

active

055388582

ABSTRACT:
A rapid radioactive method of measuring enzymatic activity of a protein kinase is disclosed. The method is an improvement to existing methodology which involves phosphorylating a peptide substrate using .sup.32 P-ATP, adsorbing the phosphorylated peptide to a solid phase, washing the phase to remove non-adsorbed .sup.32 P-ATP, and measuring the radioactivity of the phosphorylated peptide adsorbed to the phase. The disclosed improvement uses a membrane as the solid phase and positions the membrane within a chamber to separate the chamber into a first and second region. Washing is accomplished with centrifugal force; the washed solution being forced through the membrane from the first region into the second region.

REFERENCES:
patent: 4923802 (1990-05-01), Gallis
patent: 5141852 (1992-08-01), Egan et al.
patent: 5215888 (1993-06-01), Shalteil et al.
Toomik et al, Analytical Biochemistry, vol. 209, pp. 348-353, 1993.
Casnellie et al, Methods in Enzymology, vol. 200, pp. 115-120, 1991.
Sigma Catalog--Biochemicals and Organic Compounds for Research and Diagnostic Reagents, pp. 1072-1073, and pp. 854-857, 1993.

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