I am looking for an assay kit that can successfully measure extracellular ATP concentrations in vitro.
My lab currently uses an ATP assay kit that is to measure intracellular ATP, https://www.caymanchem.com/pdfs/700410.pdf
Has anyone been able to adapt this type of assay for extracellular ATP instead of Intracellular? I have tried, but it produced readings all over the place. Perhaps some components of the culture media affected the assay, and some other buffer system would be better?
Or does anyone have any other assay kit recommendations for measuring extracellular ATP in vitro?
The kit does not depend on any cell component, so it should work fine unless your culture media contains luciferase inhibitors. The critical step is to have conditions optimal for the enzyme, so using their Assay buffer is important. I wonder how your readings correspond to the kit calibration?
if you are not satisfied with the kit you are currently using, you might want to consider more established brand like promega. I used before their cell viability assay kit celltiter-glo luminescent cell viability assay that measures ATP as a detection of viability. Since your goal is to detect ATP, the same reagents should serve the same purpose. What you need to establish in addition is to have an ATP standard curve which allows you to estimate ATP concentration relative to the RLU you get from the measurement.
There seems to be a book chapter on it! Hope it is accessible and useful for you.
Article Assessment of Extracellular ATP Concentrations
Dear Maria, I think the message I just sent did not come through. If so contact me at [email protected]. I can solve your problems (cf. attached paper). Best, Arne
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