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US 9,284,597 B2Granted
Methods to assay kinase activity
AssigneeFluidigm Canada Inc.
InventorOlga Ornatsky
Life of this patent
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Abstract
Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS. The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by enzymes involved in post-translational modifications, including kinase and phosphatase enyzmes.
Claims7
A method to assay kinase activity, the method comprising:
- (a) introducing non-phosphorylated substrates with element tags into live cells;
- (b) incubating the cells having the introduced non-phosphorylated substrates with an agonist or an antagonist of kinase activity;
- (c) fixing and permeabilizing the cells;
- (d) incubating the cells with element-labeled antibodies directed against phosphorylated kinase substrates;
- (e) separating the cells from unbound antibodies;
- and
- (f) determining an effect of the agonist or the antagonist on cell kinase activity or specificity by measuring the phosphorylated substrates with element tags and attached element-labeled antibodies by using elemental analysis to detect the element tag associated with the phosphorylated substrate.
4 more elements
Dependencies are inferred from the claim text.