AAT Bioquest

How do I detect intracellular ATP levels?

Posted May 9, 2023


There are several methods you can use to detect intracellular ATP levels, some of which are: 

  1. Bioluminescence assay - Bioluminescence assay is based on the firefly luciferase enzymatic reaction, which generates photons of light using ATP from viable cells. A luminometer is used to detect the intensity of the luminescent signal. A higher luminescent signal indicates higher ATP levels.
  2. Fluorescence assay - This method utilizes a fluorescent dye, such as quinacrine or lucifer yellow, which binds to ATP and emits fluorescence. A fluorescence microscope or a plate reader is used to measure the fluorescence signal. 
  3. High-performance liquid chromatography (HPLC) - In this method, ATP is extracted from cells and tissues and separated from other components using HPLC. The amount of ATP is then quantified using UV detection or fluorescence.
Additional resources

ATP transporters in the joints


PhosphoWorks™ Luminometric ATP Assay Kit *DTT-Free*

PhosphoWorks™ Fluorimetric ADP Assay Kit *Red Fluorescence*