Help- lysing cells with triton x-100 - (May/03/2011 )
I want to lyse fibroblast to measure intracellular ATP. I saw a paper using 1% triton x-100 as lysing buffer, incubated at 37C for 30 mins in 48-well plate, and measure ATP in lysate. Could I just take the liquid from the plate using pipette after the 30min? Or do I need to scrape cells off from the plate, centrifuge and take the supernatant?
Thanks a lot!
In our experience at Promega, intracellular ATP is extremely labile in cell lysates. With our original systems, we recommend a TCA extraction to eliminate ATPases and stabilize the sample. We have ready-to-use ATP assays (CellTiter-Glo) that are much more convenient. The chemistry in this assay is designed to stop ATPases as well. In summary, Triton x-100 is NOT enough to keep the ATP from degrading and I highly recommend looking at alternative protocols.