I have some preliminary data using this method. Although the authors of the paper use GFP targetted to the ER, digitonin preserves the nuclear membrane too, so I've used nuclear-localized EGFP instead. Following the protocol described in the paper, it works great with nice CV's (not to mention the extreme ease and simplicity of this protocol). Rizwan Haq Ontario Cancer Institute Princess Margaret Hospital haqr@oci.utoronto.ca -----Original Message----- From: Ann Atzberger <Ann.Atzberger@EMBL-Heidelberg.de> To: cyto-inbox Date: Monday, February 01, 1999 5:07 PM Subject: GFP and Cell Cycle analysis NEWS > >Hallo everybody, > >In the journal Biotechniques 26:102-106 there's a report on Cell Cycle analysis >in transfected cells, using Digitonin treatment of cells instead of ethanol >fixation. Digitonin preserves the GFP fluorescence. They show CV's of >average 4% for U2OS and NIH 3T3 cells. > >Authors:Dimitri G. Pestov, Marina Polonskaia and Lester F. Lau. >University of Illinois College of Medicine. > > > >If any of you out there are interested in this and can't get the journal, >let me know and I will send the protocol. > > >Ann Atzberger. > > > >
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