Hello,
I recently did an ATP assay to detect cell viability. I used the following ATP Determination kit - https://www.thermofisher.com/order/catalog/product/A22066#/A22066.
Does anyone know the limitations of this kit?
Also, does anyone know if this kit reads other components than ATP? Are there interfering compounds that can be detected? If so, what are these compounds?
Thank you.
What kind of problem are you having? There are too many answers, you need to explain what is wrong.
I used this kit to determine cell viability after separate treatments of a chemotherapy drug and a cytoprotective agent.
Theoretically, when the ATP experiment is done properly, I am supposed to observe a greater luminescent signal at lower concentrations of these treatment compounds. At higher concentrations, these compounds present great toxicity.
However, during my ATP experiment, I did not lyse my cells. I thought that the lysis step was included in the ATP kit in question. So theoretically, I wasn't supposed to observe much luminescent signals, since no ATP was released. Yet, I obtained increasing luminescent signals when the treatment compounds were at lower concentrations. Thus, I obtained the expected results if the experiment had been done properly.
So, I don't really know what I measured. I'm trying to see if there are any compounds that could have interfered with my experiment and created luminescent signals. The solvent used for the chemotherapy drug was ddH2O and the solvent used for the cytoprotective agent was DMSO.
Dear Vicky,
it is possible that your kit components led to a limited lysis of your cells (e.g. by osmotic stress etc.), giving you the expected results. Also note that healthy cultures may leak a certain low amount of ATP e.g. by spontaneously dying cells. Also, make sure that your culture and all reagents are free from bacterial contamination, which may strongly increase the signal obtained.
Nevertheless, if you did not follow the kit's instructions, I would recommend to principially repeat the test since you cannot completely rely on the results.
Regards,
Patrick
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