facscan

[Martin Offterdinger]

In article <51bv39$ffd at mserv1.dl.ac.uk>, Patrice Chagnaud <chagnaud at infobiogen.fr> says:
>
>Hi,
>Does anybody have experiencein labelling glycoproteins with Abs
>(expressed in insect cells) and visualization by FACScan.
>thanks
>chagnaud at infobiogen.fr

Dear Patrice

Unfortunately this seems to be very difficult, 
as the baculovirus infected cells tend to be dead and permeable when 
you try to stain them with FITC-I have tried to stain for an expressed 
transmembrane protein but never really succeeded. If I had to try it again
I would keep the infection period as short as possible to have some chances
to find some non permeable cells.The problem is, if you have permeable(dead)
cells the will absorb your antibody non specifically, and you will always find
"staining" of the cells.I would also try to get the cells into suspension very 
carefully, this might increase your chances to see some specific binding.
What about trying cellELISA instead of FACS.
Bye Martin