Nuclear extracts from frozen adipose tissue - (May/14/2013 )
I have frozen adipose tissue; not possible to generate new, fresh tissues. I would like to do an EMSA to screen for transcription factors involved in reduced transcription of a gene of interest, which usually requires preparing nuclear extracts. Is it possible to make nuclear extracts from frozen adipose tissue? Can I bypass the nuclear extract prep? If not, can you suggest an alternative approach? I also have limited paraffin-embedded tissue available.
Most of the time, freezing tissues and then thawing them results in damage to the membranes, including the nuclear membrane, so you are likely to have problems if you do decide to go ahead. Having said that, it depends on how the tissues were frozen, if they are in something that prevents the formation of ice crystals, like OCT or DMSO, then you may have a shot at getting intact tissue out.
Can't comment on the bypassing the nuclear prep without knowing exactly what it is you are looking for (and even then may not know).