Has anyone tried assaying for luciferase activity on previously stored but not lysed cell pellets? The cells were scraped in PBS and spun down and the pellets were stored at -80. If the pellets are sonicated in PLB buffer, do you still expect to detect luciferase activity?
Dipsikha hi,
Most types of luciferase are quite stable (except for short live/destabilized versions of Luciferase), so if you stored your cells immediately after scraping @ -80C they should retain the Luciferase activity.
With that said you should not rely on the results quantitatively, but rather quantitatively, hence yes/no and maybe low/high.
If you need quantitative results and analysis, you'd better use fresh cultures and prepare extract to be tested, preferably soon after their preparation, with the appropriate positive & negative controls, and replicates.
Best
G
Thank you @Ghil Jona and @Ajay for your inputs. I will definitely keep this in mind.
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