EGFP

Deborah Berglund umbbd at gemini.oscs.montana.edu
Mon Aug 24 12:57:03 EST 1998


You may be causing the cells to leak with your trypsin treatment.  Can you
get the cells off by incubating (10-15 min) them with two or three changes
of Calcium and Magnesium free PBS (we use Tyrode's), then rinsing them off
with pasteur pipets,  gently.
Deb

On Mon, 24 Aug 1998, Jacqueline Saleh wrote:

> 
> Dear Flow-ers:
> 
> We are working quite a lot with EGFP and are seeing quite a discrepency in
> results
> between what we see under the scope and the fluorescence readout on the
> instrument.
> I looked at two samples under the fluorescent scope and the difference in
> fluorescence is
> quite significant. Cells are then trypsinized, washed and read using the
> FacsCalibur and
> the results of these two samples are quite similar. I thought maybe the
> trypsin might be
> pulling off the EGFP but EGFP is internal. Does anyone know what might be
> going
> on here????? Thanks in advance for all your help.
> 
> Jackie Saleh
> Ariad Pharmaceuticals
> saleh at ariad.com
> 
> 




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