Is it ok and possible to remove the supernatant (cell media) containg secreted luciferase and freeze in a 96 well plate at -20 or -80 to thaw later for luciferase assay. I am using a secreted Nanoluc reporter vector.
Thanks
For better results, proceed for the assay immediately after you collect the media. If you want to freeze for short term -20 or for long term -80 is fine but, freeze thaw cycles reduce activity of any enzymes including luciferase.
As Goodwin has suggested, it always better to process the samples early or fresh, however, if you have limitations, then avoid repeated freeze thaw cycles. Good Luck
Half-life of the secNLuc enzyme is 7 days at 37 degrees, so for short term storage it is not necessary - just store at 4 degrees. If you are collecting samples over time in order to run the activity assays simultaneously, sure - toss them in the freezer, they'll be fine. Just be sure to treat all the samples in the same manner (e.g. freeze all samples, including the last). Be sure to vortex briefly or pipette to thoroughly mix the thawed solutions as they'll stratify. The enzyme is so much more sensitive compared to firefly & renilla that you won't notice any difference in activity. I do this routinely.
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