ATP cell viability assay

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
TK6 and HepaRG cell line
Used species
Human cell line


Method keywords
  • cell viability test
  • in vitro
  • ATP
Scientific area keywords
  • in vitro cytotoxicity
Method description

The ATP cell viability assay (CellTiter-Glo assay test kit) provides a quantitative measurement of the ATP content directly proportional to the number of cells present in culture. The homogeneous assay procedure involves adding one single reagent, CellTiter-Glo Reagent directly to cells cultured in serum-supplemented medium. This results in cell lysis and generation of a luminescent signal proportional to the amount of ATP present. Luminescence can be measured already 10 minutes after adding reagent & mixing. The half-life of the luminescent signal is greater than five hours so the measurement can be performed even hours after adding the reagent. The CellTiter-Glo® Assay relies on the properties of a proprietary thermostable luciferase (Ultra-Glo™ Recombinant Luciferase), which generates a stable “glow-type” luminescent signal. % Cell viability after exposure is calculated through comparison of measured RLU of unexposed control cells (set to 100% viability) and exposed cells.

Lab equipment

Standard equipment for working with cell cultures, luminometer

Method status
  • History of use
  • Published in peer reviewed journal

Pros, cons & Future potential

  • Sensitive (even few cells can be detected),
  • Fast (data available 10 minutes after adding reagent),
  • Easy,
  • Very stable luminescent signal flexible (ATP content from both adherent and suspension cells in different well formats can be measured),
  • No interference with compounds with oxido-reductive potential (MTT) or colored compounds (NRU).
  • Expensive (test kit),
  • Sensitivity can be a disadvantage (the occurrence of metabolic oscillations can cause fluctuations of the ATP content per cells).

References, associated documents and other information


J Shi, S Springer, P Escobar (2010) Coupling cytotoxicity biomarkers with DNA damage assessment in TK6 human lymphoblast cells 10.1016/j.mrgentox.2010.01.008

Associated documents
CellTiterGlo Luminescent Cell Viability Assay TB288.pdf


Chemical and physical health risks