For those of you who are new to the list it might prove useful if you search the archives @ http://flowcyt.cyto.purdue.edu/ before posting questions to the listserv. Margaret >Hi All, > I agree with you, Derek. I will not answer questions where the person >obviously has done no work at all. The discussion on replys has taken place >before. I understood that the questioner would place a synopsis on the >board. Requesting a list from the questioner sounds like a good idea as well. > >Paula > >At 12:20 PM 3/15/2001 -0500, Derek Schulze wrote: > >>Two things here: >> >>You are more likely to get responses to your question if you ask more >>specific questions. If you want to work with Annexin V first go to the >>literature and see what has been done with your cell type. >> >>Some key things to think of to begin with though: >> >>-AV works best with non adherent cell lines because trypsinizing can expose >>PS on the outer plasma membrane leaflet giving false positives. >>-think of a good positive control that is appropriate for your cells >>(ceramide treatment, serum starvation or anti-FAS etc...) >>-optimize your concentration and exposure time for whatever method if >>inducement you are going to use so as to minimize necrosis >>-if it is appropriate confirm your outcomes with a fluorescence microscope >>(only really good for later apoptosis - nuclear fragmentation) >>-AV is generally used for very early apoptosis signals, for more precises >>results you might consider doing a tunnel assay (tdt labelling of DNA >>fragments with BrdU-FITC) for slightly later apoptosis events. >>----------------------------------------------- >> >>People not responding directly to questions may have to do with the fact >>that when the reply button is hit the email from the postee is presented >>NOT the mailing list. This should be changed so that the mailing list is >>presented first. I have to hit "reply to all" in order to get the mailing >>list address to show up. Maybe this has something to do with how I have >>Eudora set up but I can't see anything obvious. >> >>> > On Mon, 12 Mar 2001, Ashman, Margarita wrote: >>> > >>> > > >>> > > Does anyone have experience with annexin V for running apoptosis >>> > > >>> > > Thanks >>> > > >>> > > Margarita Ashman >>> > > mashman@med.miami.edu >>> > > >>> > >> >>Derek Schulze >> >>Cancer Research Labs >>Queen's University >>353 Botterell Hall >>Kingston, ON, K7L 3N6 >>(613) 533-6635 >> >>http://meds.queensu.ca/medicine/crl/ > > >Paula Fukushima >Flow Cytometry >LP, NCI, NIH >10 Center Drive MSC-1500 >Bethesda, Md. 20892-1500 >>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>> Margaret M. Tropea Critical Care Medicine Department National Institutes of Health (ph) 301-496-7752 (fax) 301-480-3389 >>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>
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