Extraction of nuclei from frozen tissue
Katie Ridd
Monday, 29 June 2009 09:08 UTC
Note from moderators: the following query was posted in response to another forum thread.
We have re-posted it here to increase its visibility.
Hi all!
My name is Stella and I am a PhD student in Germany, working in a proteomics department. I got one question that I could not solve alone til now and I am wondering if anyone from you can help me. I must analyse nucleus extracts from liver tissues but in our Institute the only SOP that explained that forbids to use tissues already frozen. Do anyone from you know an alternative protocol for that? (My samples are at -80C)!
I thank you all in advance,
sincerely,
Stella
Updated 29 June 2009 09:13 UTC
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Replies
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In the past I have worked with cells not tissue but…. I would assume you thaw the cells/tissue on ice and follow standard procedures, such as those described in Current protocols in “cell biology” or “molecular biology” to extract proteins. Typically it is assumed that protein extracts from frozen samples are less biologically active than those from fresh samples. If you have no option but to use frozen cells/tissue then why not run controls using fresh cells/tissue at the same time and devise a positive control common to both samples?
Good luck -
Anonymous
Thanks Mark! Well, actually I was just wondering if maybe the nucleus can be broken from freezing… I have found not so much about that….
Anyway, I will run even the extracts from the fresh tissue, if it is possible and I will see! Thanks for the tip
;)
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