I'm inducing M0 macrophages from THP-1 by using Phorbol 12-myristate 13-acetate (PMA)
But frome time to time, I encounter induction failure (twice now), which could be fixed by new PMA stock.
I would like to have some suggestions on storing PMA?
Currently I preserve 5000x and 1000x stock,
5000x stock dissolved in DMSO, and 1000x was diluted from 5000x with PBS.
All stock was store in -20°C.
Is there any suggestions about how many freeze-defreeze cycle PMA could encounter, or if PMA can't left on ice for too long?
Thanks!
Hello Yuhung Zheng
Here are some suggestions.
You make a stock of 5000x in DMSO and then make aliquots of this stock and store in -20 degree C under dark conditions. This stock will last for least 6 months. Make aliquots of this stock in such a way that it could be used only for single use so that repeated freeze-thaw cycles could be avoided.
Do not preserve 1000x diluted stock made in PBS. You will have to make fresh diluted stock in aqueous medium each time you perform your experiment. Such diluted stocks are not stable when prepared and stored.
The diluted stock and the working solution of PMA should be prepared and used on the day of the experiment and could be reused on the same day if you are planning for another experiment on that day. Do not store the diluted stock and working solutions of PMA.
Best Wishes.
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