Principle of ATP measurements
ATP participates in the oxidization of luciferin by the enzyme firefly luciferase, a reaction that produces light. The quantity of light produced is proportional to the quantity of luciferin oxidized. This reaction is very popular in reporter gene assays, where ATP and luciferin are in excess, and the quantity or light produced is limited by the expression of luciferase (the reporter). However, the same reaction can be used to quantify ATP if both luciferin and luciferase are in excess.
Applications of ATP measurements
ATP measurements are used to monitor raw materials, manufacturing plant or food, drugs and healthcare equipment for bacterial contamination, often for hygiene monitoring, as well as for waste water analysis.
In the biotechnology and pharmaceutical industries, ATP measurements are used to evaluate cell proliferation, apoptosis and cytotoxicity. Cell viability assays, e.g. in tumour chemosensitivity assays and antibiotic susceptibility testing and determination of enzymes and metabolites in ATP-linked reactions can be performed. The bioluminescent assay of ATP has become very popular because of its high sensitivity and convenience.
Application Notes related to ATP measurements
ATP measurements with the Orion II Comparison of standard curves for ATP with the Orion II Microplate Luminometer
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ToxiLight™ Cytotoxicity Assay with the Orion II ToxiLight™ Non-Destructive Cytotoxicity Bio Assay Kit (Lonza) with the Orion II Microplate Luminometer
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ViaLight™ Assay with the Orion II ViaLight™ Cell proliferation and Cytotoxicity BioAssay Kit (Lonza) with the Orion II Microplate Luminometer
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Rapid Bacterial Detection with the Sirius L Rapid Bacterial Detection Assay (Useful Biology) with the Sirius L Tube Luminometer
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ATP detection limit with Sirius L Below amol ATP detection limit with Sirius L Tube Luminometer and ATP Reagent SS from BioThema
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Recommended Instruments for ATP measurements
The second most important element needed for the quantification of ATP is a luminescence reader. Hand-held luminometers are popular for hygiene monitoring of surfaces. However, its sensitivity and dynamic range are very limited, and normally suitable only for qualitative analysis. For assays requiring high sensitivity and reliability, a high-performance laboratory instrument is recommended; options include tube luminometers and luminescence microplate readers.