Background

Adensoine triphosphate (ATP) is the organic chemical that cells use to transport the energy they need to drive metabolic processes.
Measurement of total ATP content is useful as an indicator of the overall viability of cell models
ATP is useful in the context of plate reader based assays to calculate IC50 values, or to identify potentially toxic compounds in a high throughput manner.
Using Celltiter-Glo, ATP content can be assessed in 3D cell models to evaluate overall viability of cells within the model.
This assay can also be conducted on adherent monolayers of cells.
A related assay, Realtime-Glo, looks at the metabolic redox potential of cells. Realtime-Glo is useful as it is not an endpoint assay, allowing for the monitoring of cell model viability multiplexed with imaging based endpoints. Using Realtime-Glo, counter screens for toxicity can be conducted on the same disease models being tested for compound efficacy, or be used to triage models for more in depth imaging for mechanisms of cell death or dysfunction.

Protocol

Instrument	ThermoFisher Varioskan
Analysis Method	Plate Reading Luminometer
Markers	Luminescence based Viability Luminescence based Metabolic Potential
Cell Types Available	ATCC cell lines (e.g. HepG2, A549, BT549, MCF-7, and others) Other cell lines available on request
Test Article Concentration	8 point assay (0.05, 0.1, 0.5, 1, 5, 10, 50, 100 µM) (custom concentrations available)
Number of Replicates	3 replicates per concentration
Quality Controls	0.5% DMSO (vehicle control) Staurosporine (positive control)
Test Article Requirements	50 uL of 20 mM solution or equivalent amount of solid
Data Delivery	Dose response curves, IC₅₀ values

Multiplexing Alternative