Luciferase assay for determining the physical condition of cells
A method for determining the physical condition of cells comprises (a) adding an agent to cells which releases adenylate nucleotides therefrom, for example an anionic or non-ionic detergent; (b) adding luciferin and luciferase and accordingly determining the ATP concentration by measuring the intens...
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Main Authors | , , |
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Format | Patent |
Language | English |
Published |
16.09.1998
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Edition | 6 |
Subjects | |
Online Access | Get full text |
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Summary: | A method for determining the physical condition of cells comprises (a) adding an agent to cells which releases adenylate nucleotides therefrom, for example an anionic or non-ionic detergent; (b) adding luciferin and luciferase and accordingly determining the ATP concentration by measuring the intensity of light emitted; (c) when ATP levels are substantially constant, adding a reagent which converts ADP to ATP, preferably a mixture of phosphoenol kinase and pyruvate kinase, and measuring the light emitted to determine the ADP concentration; (d) determining the ADP:ATP ratio. The method can be used to distinguish between, apoptotic cells, necrotic cells, proliferating cells and growth arrested cells. The magnitude of the ADP:ATP ratio will be necrotic cells > apoptotic cells > proliferating cells > growth arrested cells. The application of the method to mammalian cell lines and a kit for carrying out the method are described. The anti-cancer drugs camptothecin and etoposide are incorporated into some of the assays. |
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Bibliography: | Application Number: GB19970027296 |