I want to set up an assay for Apoptosis and have ready my control with a "X" % positive events for Annexin V and DAPI.
Can I pre-treat cells with Staurosporine, and then freeze them and keep them at LNF as controls with a known % of positive events? will that work when I will thaw them or all the apoptotic cells will dye during the process of freezing?
did anyone try it already?
thanks
I have not tried it, but doing cell culture for long time. And there is definitely loss of cells when you thaw a freezed cells. Recovery does not more than 60% practically.
So your X% cells will be around half of X%.
You should perform this exercise Everytime.
I wouldn't do this. The freeze thaw will be too harsh on the cells and your predetermined % viable number will be inaccurate.
If you want a comparator for the experiment, you should be running the staurosporine condition in parallel to your experimental samples.
I have not tried it, but doing cell culture for long time. And there is definitely loss of cells when you thaw a freezed cells. Recovery does not more than 60% practically.
So your X% cells will be around half of X%.
You should perform this exercise Everytime.
Hi Ana,
I wouldnt recommend freezing and thawing of staurosporine treated cells. It will definitely lead to cell death, which will interfere with the experimental data.
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