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AAT Bioquest

How is luciferase used to detect ATP?

Posted May 12, 2021


Answer

ATP plays an important role in energy exchange in biological systems. A lot of methods have been used for ATP determination. The most successful technique is the bioluminescent method. Luciferin is catalytically oxidized by the enzyme luciferase in a high quantum yield chemiluminescent reaction. Under optimum conditions, light intensity is lineary related to ATP concentration. ATP can be monitored by direct lysis of the cells with suitable detergent, the released ATP is then free to react with the luciferin-luciferase and leading to light emission at 562 nm. A luminometer can be used to measure the light intensity.

Additional resources

Use of luciferase probes to measure ATP in living cells and animals

PhosphoWorks™ Colorimetric ATP Assay Kit