Hello Sunetra, I have used Sytox Green for cell cycle analysis and the fixation protocol was the same as for PI staining ie fix in cold 70% ethanol for at least 30mins. I think it would also be OK with a detergent permeabilisation as well. You don't say at which concentration you used the dye. I was using it at 1uM but it may be that you might have to try a range. Also it is often a good idea to leave the samples to stand for a while (note that scientific term) before running as staining may some time to get to equilibration. I found comparable CVs to PI staining which I was using as my standard. Good luck! Derek >Hi >I was wondering if anybody has ever done cell cycle analysis with >Sytox green (molecular probes) dye. And if so, can you please >forward me a protocol. >I tried the same procedure as normally used with Propidium iodide, >but I couldn't see the normal cell cycle pattern. I saw a very broad >peak (probably G1), but nothing else. >Any help will be highly appreciated. >Thanks, >Sunetra -- *************************************************************** Derek Davies Voice: (44) 020 7269 3394 FACS Laboratory, FAX: (44) 020 7269 3479 London Research Institute, e_mail: derek.davies@cancer.org.uk Cancer Research UK mobile: 07790 604112 44 Lincolns Inn Fields, London, UK. Web Page: http://science.cancerresearchuk.org/sci/facs/ In tenebris lux ***************************************************************Received on Fri Mar 11 17:18:00 2005
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