Question (76043) | Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) (ab109721)

Go to datasheet (ab109721)

Question

I'd like to use the tissue, once processed using ab110169, for the following kits 1) ab109721, 2) ab110413 and ab109902.
So the main question is, can I store the brains by freezing (eg. liquid nitrogen to -80 deg C) and thaw the brains and produce extracts that will produce viable sample for use with all 3 kits noted above.

Answer

Yes, you can extract mitos from frozen tissues but be aware that freezen/ thaw cycle would weaken the cell membrane and cause less recovery of intact mitochondria.

A: There will be slightly loss if you extract enzyme from frozen mitos versus fresh mitos due to the degradation. Adding protease inhibitor to your mito prep would minimize the loss.

As long as the tissues are properly frozen, the variant between different time points should be minimum (within weeks). However, multiple freezen/thaw cycle will cause difference. If you plan to extract sample from different days, make sure you make aliquots of your samples. if you plan to compare the enzyme activity from different samples, we highly recommend to prep the samples under the same condition.

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