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Features of the ATP Determination Kit include:
Principle of the ATP-dependent bioluminescent reaction
The ATP Determination Kit is based on the absolute requirement of luciferase for ATP to produce light. In the presence of Mg2+, the luciferase enzyme catalyzes the reaction of luciferin, ATP, and O2 to form oxyluciferin, AMP, CO2, pyrophosphate, and light with an emission of approximately 560 nm at pH 7.8. The luminescent signal generated is directly proportional to the amount of ATP, allowing for the measurement of ATP levels.
Measure ATP levels in solution or cell lysates
The assay reagents in the ATP Determination Kit are not permeable to live and intact cells, making this assay suitable for measuring extracellular ATP or ATP from samples in solution. To measure intracellular ATP levels for cell health or viability assays, cell lysis is required to release ATP from the cells. Lysis buffer options that are compatible with the luciferase-luciferin reaction include the Pierce Luciferase Cell Lysis Buffer (2X) (Cat. No. 16189) and the 20X cell lysis buffer formulation listed below. The 20X buffer should be diluted 1:20 with water and used immediately for cell lysis prior to running the luciferase ATP assay.
20X lysis buffer:
Comprehensive and versatile ATP assay kit
ATP is a critical molecule found in all living cells and is the energy source for various biological processes, including muscle contraction, nerve impulse propagation, and biochemical synthesis. ATP levels are often measured to determine cellular viability and metabolism, assess mitochondria function and activity, or detect bacterial and microbial contamination. This luciferase-based ATP detection kit provides a versatile and sensitive method for measuring ATP production in various enzymatic reactions, including ATPase and NADPH oxidase, and monitoring ATP release from cells.
The luciferin–luciferase luminescence assay can be used to detect low-level bacterial contamination in biological samples such as blood, milk, and urine, along with environmental samples for testing of water, soil, and sludge. ATP detection has also been used to study the effects of antibiotics on bacterial populations, determine cell proliferation and cytotoxicity in both bacterial and mammalian cells, and distinguish cytostatic versus cytocidal potential of anticancer drugs on malignant cell growth.