Components

One vial of Luciferase ATP Monitoring Enzyme, Enzyme Reconstitution Buffer, 1 bottle of Nucleotide Releasing Reagent, one vial of ATP Standard, and a user protocol.

General description

The assay is based on the firefly luciferase-catalyzed oxidation of D-luciferin in the presence of ATP and oxygen, whereby the amount of ATP is quantified by the amount of light (hv) produced.

Cell death (especially apoptosis) is an energy-dependent process that requires ATP. As ATP levels fall to a point where the cell can no longer perform basic metabolic functions, the cell will die. A typical apoptotic cell exhibits a significant decrease in ATP level. Therefore, loss of ATP level in the cell has been used as an indicator of cell death. In contrast, increased levels of ATP has been an indicator of cell proliferation. Calbiochem^® brand ATP Assay Kit utilizes bioluminescent detection of the ATP levels for a rapid screening of apoptosis and cell proliferation simultaneously in mammalian cells. The assay utilizes luciferase to catalyze the formation of light from ATP and luciferin, and the light can be measured using a luminometer or Beta Counter.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Packaging

1 kit in Glass bottle

Preparation Note

Reagent Reconstitution1. Reconstitute ATP Monitoring Enzyme with 2ml of the Enzyme Reconstitution Buffer. Mix well by gentle pipetting. The reconstituted enzyme is stable for up to 2 months at 4°C.2. Prepare an ATP standard solution by dissolving the 1 mg ATP into 1 ml of H₂O. The solution is stable for several weeks at -20°C.

Principle

The assay can be fully automatic for high throughput (10 s/sample) and is extremely sensitive (detects 10-100 mammalian cells/well). The high sensitivity of this assay has led to many other applications for detecting ATP production in various enzymatic reactions, as well as for detecting low level bacterial contamination in samples such as blood, milk, urine, soil, and sludge.

Specifications

Assay Time: 1 h

Storage and Stability

Upon arrival store the entire contents of the kit at -70°C.

Warning

Toxicity: Multiple Toxicity Values, refer to MSDS (O)