high background with FDG for beta-gal FACs analysis

From: Hadi (Hadi@pob.huji.ac.il)
Date: Wed Feb 28 2001 - 11:30:28 EST


Dear All:

I used the FDG for FACs as follows:
> I injected murine stem cells marked with the bacterial beta-galactosidase
> reporter gene intravenously to syngenic C3H/HeN female mice and then
> 24hours post-injection i collected the cells from whole BM and spleen,
> stained them with FDG and analysed them with FACs to estimate the
> percentage of cells expressing b-gal, but unfourtinately i had very high
> background of unspecificly FDG-stained cells (30-40% of sorted cells in
> both organs) although i used the chloroquine reagent to inhibit endogenous
> b-gal.
> As you see, i'm trying to detect very low events, so high background is
> very disturbing.
> If any body had experienced similar results or have any idea to decrease
> the background i would appreciate his feed back..
> Thanks,
> Hadi

+--------------------------------------------------
Hadi Aslan
Msc./D.M.D. student
Laboratoy of Molecular Pathology-Prof. Dan Gazit
School of Dentistry-Hadassa Ein-karem
Hebrew University of Jerusalem
Cellular : 051-450259
Lab Tel: 02-6757625
+--------------------------------------------------



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