Immunofluoresence problems

Fergus Doherty Fergus.Doherty at nottingham.ac.uk
Fri Feb 6 06:15:31 EST 1998


I have a S. cerevisiae strain expressing a c-myc epitope tagged protein. 
Following cell wall removal and permeablisation with organic solvents I can
localise the protein by immunoperoxidase  (HRP anti-mouse), but using a
SIGMA FITC anti-mouse the signal is poor.  Any clues as to how to improve
the signal?  I find I get high backgrounds with the avidin-biotin system
(SIGMA FITC-Avidin), so that isn't an option.

I could just stick with immunoperoxidase, but then I can't double label. 
Anyone know of a *visible light* alternative to DAPI to stain nuclei?

Thanks

-- 
Fergus Doherty,
Biomedical Sciences,
Nottingham University,

Fergus.Doherty at nottingham.ac.uk
0115 970 9366 (74-41366 internal)




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