Aug 09, 2023
Analyzing cellular ATP levels
- 1Lazarou Lab, WEHI
Protocol Citation: Louise Uoselis 2023. Analyzing cellular ATP levels. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp3jmjvzp/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 09, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 86213
Keywords: ASAPCRN
Abstract
Protocol for analysis of cellular ATP levels using the Promega Mitochondrial ToxGlo Kit.
Day 1
Day 1
Seed cells into white opaque walled 96 well plates in 80 µL of media/well, aiming for a confluency of ~80-90% the next day at the time of treatment. Fill all surrounding wells with water to prevent evaporation of the experimental samples. Make sure you seed a DMSO treatment control for each sample collection point.
Day 2
Day 2
5m
5m
Treat cells as desired in 80 µL of media/well.
If analysing cells cultured in galactose media, change the media to galactose media 12 – 24 h prior to sample collection.
To analyse ATP levels, warm the 2x ATP detection reagent to Room temperature , add 80 µL of the 2x ATP detection reagent to each well using a multichannel pipette
Place the plate on a shaker block set to Room temperature , cover the plate in foil, and incubate the plate shaking at 400 rpm for00:05:00
5m
Immediately take the plate to a plate reader that can read luminescence signal.