I'm trying to get subcellular fractions (specifically, I only need the nuclear fraction) from neuronal tissue. However, all my current samples were flash frozen and stored in -80C subsequently (for whole cell lysate, which was my original intent for the samples).
Does subcellular fractionation for nuclear protein require fresh tissues and cells? Will flash freezing might cause ice crystals to form and pierce the nuclear membrane, so I wouldn't get good separation?
Thank you in advance.
Dear Amy, do you want to look at it (Electron microscopy) or actually work with them? The latter, I am afraid, won't be possible. With best wishes, Matthias
Dear Amy,
I don't know for nuclear t fraction but for other subcellular fractions pre or post membrane, you can perform it following flash frozen and _80 storage?
Eric
it MAY BE POSSIBLE BUT IT WILL DPEEND ON YOUR SPECIFC SITUATION AND WHAT YUO ARE TRYING TO ISOLATE - ICE CRYSTALS DO PIERCE THE NUCLEAR MEMBRANE
I would try to use fresh tissue. Why flash freeze tissue. Start the day very early using fresh tissue and try to isolate organelles.
yes - I'd agree if its possible use fresh - leaves out any unexpected variable - good luck
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