Adenosine 5 triphosphate atp bioluminescent somatic cell assay kit

The Adenosine 5′-triphosphate (ATP) bioluminescent somatic cell assay kit (Sigma-Aldrich) was used to determine cell viability, as described by the manufacturer. Briefly, cells in each well of a 96-well plate were lysed in 45 µl of the Somatic Cell ATP Releasing Reagent. Of this volume of lysate, 30 µl was transferred to a white-walled 96-well plate. To this was added 30 µl of resuspended Adenosine 5′triphosphate (ATP) Assay Mix and the resulting luminescence measured using the Orion II Microplate Luminometer (Berthold Detection Systems). To calculate Relative IC₅₀ values, the resulting data were fitted to a Sigmoidal Dose-Response Curve (variable slope), as implemented in GraphPad Prism. The TOP parameter was fixed at 100% cell viability.

Single oocyte ATP content was determined using an Adenosine 5′-triphosphate (ATP) bioluminescent somatic cell assay kit (Sigma-Aldrich), according to the manufacturer’s instructions. The luminescence intensity was measured using a Luminometer (Promega GloMax® 20/20).