Cellular ATP

Cellular ATP Kit HTS is intended for enumeration of viable cells by quantification of cellular ATP (adenosine triphosphate). The assay is intended for rapid analysis of cytotoxicity and proliferation in cell studies. The assay of cellular ATP replaces methods such as tritiated thymidine incorporation and tetrazolium salt reduction. If a stable light emission is not obtained, the ATPases are not completely inhibited. We then recommend the Cell Viability Kit SL containing a strongly ATPase inactivating agent.

The Cellular ATP Kit HTS is especially designed for HTS screening methodology; enabling assay in 96 well format with manual or fully automated handling. The assay format is a mix and measure addition of ATP Reagent containing cellular lysis agent and stabilizers for the firefly luciferase enzymatic reaction. The T1/2 of the light emission is 3 hours in medium containing 10% FCS (Fetal Calf Serum) and 12.5 hours in media supplemented with e.g. vitamin mixture F12, enabling screening of a large number of samples. The Cellular ATP Kit HTS is useful for quantification of ATP in range of 10^-11 – 10^-7 mol/L. This dynamic range covers the commonly used seeding cell number and culturing media.

In samples with a high density of ATPases, e.g. nerve cells, the ATPase inhibitor in the Cellular ATP Kit might not completly inhibit the ATPases. This will result in a slight decay of light. In such cases, a luminometer with dispensers could be used. Alternatively use 188-441 Cell Viability Kit SL which contains a stronger ATPase inhibitor.

The stable light makes it possible to manually add the ATP Reagent and the ATP Standard with a multi-channel pipette or to use a microplate luminometer with dispensers.

Detailed described procedures are found in the “Instructions for Use”, which is included with every kit.

Number of assays:

1200 tests depending on whether assays are performed in microplates.

Detection limit:

Quantitative detection of ATP in the range of 10^–11 to 10^–7 mol/L.