solubilizing nuclei

Martin Offterdinger a8803349.nospam at unet.univie.ac.at
Mon Apr 6 04:44:45 EST 1998


On 5 Apr 1998 15:00:15 +0100, "Wolfgang Schechinger"
<wgschech at med.uni-tuebingen.de> wrote:

>Hi all. 
>
>I've got dishes in the freezer where I had treated cell layers with 
>triton-x-100 for solubilizing plasma membranes and cytosol. 
>Now the nuclei are frozen down there and I'm interested in measuring 
>nuclear enzyme activity (WHAT a *nuclear* energy power this will 
>unleash!!! ;-) So, SDS will be too heavy for that purpose (of course 
>good enough for going onto a PAGE gel with them).
>What do you suggest instead of scraping them off and sonicate 
>(SONYcate ?? - using my walkman?? - uuh sorry, that's the sunday lab 
>blues) them?
>
>Any input is welcome!
>
What about using a dounce homogeniser in hypertonic (600mM KCl) buffer
and prepare nuclear extracts?
Martin
Martin Offterdinger
Internal Med.I,Dept. Oncology
University of Vienna
Austria
E-Mail:a8803349.nospam at unet.univie.ac.at
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