Ulrich Huth <ulrich.huth at pharmazie.uni-freiburg.de> wrote:
> I want to buy Hoechst 33342 to stain my living COS-7 cells. (DAPI staining
> does not work).
> Is there anybody having experience with this dye? Molecular Probes does not
> provide a protocol for nuclei staining. Which concentrations do I use? What is
> the incubation time?
H33342 will stain your cells DNA right enough. Do you just want to use
it as a nuclear marker or quantitaively in a flow cytometer? If you just
want to use it as a general nuclear marker, I would use 1ug/ml for about
20mins at 37C. For flow cytometric DNA analysis, I would suggest
starting at 10ug/ml for 30 mins but you may need to detemine the best
staining concentration and time empirically as cell types differ (and
mdr phenotype cells will actively pump it out so can be problematic).
> Any problems or tricks with this fluorescent marker? Do you think the
> Hoechst Dye is a good marker?
Good as a quantiative DNA stain and a lovely blue under the fluorecence
The ageing jester who won't gracefully retire.