Michelle,
I have used anti-fluorescein antibodies to quench externally bound
FITC. It was a long time ago and I think I used a polyclonal
antibody. Molecular probes may be able to help you with this one.
Larry Kahn
Searle Discovery Research
St. Louis, Mo
lekahn@ccmail.monsanto.com
______________________________ Reply Separator _________________________________
Subject: Detection of FITC oligos
Author: owner-cyto-sendout@flowcyt.cyto.purdue.edu at INTERNET
Date: 6/5/97 4:34 PM
I have a query regarding detection of fluoresceinated oligos
intracellularly by FACS. Is it possible to distinguish between
externally bound liposome/oligo complexes and internalised intracellular
complexes by FACS following transfection of mammalian cells? I know
that I could use a microscope (heaven forbid!) but the number of samples
and the need for quantitation w.r.t. transfection efficiency preclude
that option (plus I'm very attached to my FACSort so would always choose
that over a microscope if I had a choice). Maybe there is a way to strip
off non-internalised oligos so you can be sure you're measuring only
internal fluorescence? Any tips from users who have tried this would be
very much appreciated.
Thanks in advance,
Michelle Miller
Johnson and Johnson Research
GPO Box 3331, Sydney NSW 2001
Australia
Ph 61 02 9360 9377
Fax 61 02 9360 9813
Email jjph13mm@angis.su.oz.au
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:49 EST