For what it's owrth, we always thaw and then sonicate in 2x sample buffer,
then boil for 10 minutes and load. Of course, it may depend on the tissue you
are working with, but for ovaries that seems to work.
Samuel Gray wrote:
> Does anyone have a good protocol for extracting total protein (for western
> blotting) from tissues frozen in liquid nitrogen? Should I pulverise the
> tissue whilst still frozen (as I would for RNA extraction) or allow the
> tissues to defrost and mince/sonicate in a protein extraction buffer?
>> Thanks for any suggestions,
>> Samuel Gray.
> Samuel Gray
> Division of Vascular Medicine,
> Derbyshire Royal Infirmary,
> DE1 2QY
> Tel: 01332 254966
> Fax: 01332 254968
>samuel.gray at nottingham.ac.uk
Brian D. Cohen, Ph.D.
Laboratory of Reproductive and Metabolic Disorders
David Axelrod Institute #5031
NYS Dept. of Health
P.O. Box 22002
Albany, NY 12008-2002
cohen at wadsworth.org
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