Methods and kits for detecting protein kinases
AssigneeLumitech (UK) Limited
InventorsSharon Patricia Mary Crouch, Kevin John Slater
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Abstract
A method for measuring protein kinase activity comprising: (a) providing a first solution comprising ATP and a protein kinase to be tested, and a second solution comprising ATP in the absence of said kinase to be tested; (b) adding a substrate capable of being phosphorylated by the protein kinase to be tested to the first and second solutions of step (a); (c) measuring the concentration of ATP and/or ADP, or the rate of change thereof with respect to time, in each of the reaction mixtures formed in step (b) using a bioluminescence reaction; and (d) using the information about the concentration of ATP and/or ADP to determine the activity of the protein kinase to be tested.
Claims42
A method for measuring protein kinase activity, said method comprising:
- (a) providing a first solution comprising ATP and a protein kinase to be tested, and a second solution comprising ATP in the absence of said kinase to be tested;
- (b) adding a substrate capable of being phosphorylated by the protein kinase to be tested to the first and second solutions of step (a);
- (c) measuring the concentration of ATP and/or ADP, or the rate of change thereof with respect to time, in each of the reaction mixtures formed in step (b) using a bioluminescence reaction;
- and
- (d) using information about the concentration of ATP and/or ADP to determine the activity of the protein kinase to be tested.
A method for identifying a compound which modulates the activity of a protein kinase, said method comprising:
- (a) providing a first solution comprising ATP, a protein kinase and a compound to be tested, and a second solution comprising ATP and the protein kinase in the absence of said compound to be tested;
- (b) adding a substrate capable of being phosphorylated by the protein kinase to the first and second solutions of step (a);
- (c) measuring the concentration of ATP and/or ADP, or the rate of change thereof with respect to time, in each of the reaction mixtures formed in step (b) using a bioluminescence reaction;
- (d) using information about the concentration of ATP and/or ADP to determine the activity of the protein kinase is the first and second solutions;
- (e) comparing the activity of the protein kinase in the first solution with the activity of the protein kinase in the second solution to identify compounds which modulate a protein kinase, whereby the compound to be tested is identified as a protein kinase modulator if the activity of the protein kinase in the first solution is different from the activity of the protein kinase in the second solution.
Dependencies are inferred from the claim text.