Apoptosis -- no direct responses

Margaret Tropea mtropea at nih.gov
Mon Mar 19 12:06:10 EST 2001


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 at 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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