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
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