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If you use methanol fixation you have to use an
anti-actin antibody to visualize actin (phalloidin
won't work). The pan-actin clone C4 works great, you
can't really see the difference with phalloidin. You
can get it from different companies. Recently tested:
the one from Accurate Biochemicals is great (1:80
dilution); we've had trouble with the one from
Chemicon since they have changed the conditioning
(even though they claim nothing has changed).

maryse


--- Marylou Zuzarte <[log in to unmask]>
wrote: > Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> I would like some opinion over the fixation of rat
> heart myocytes. I have
> tried 4% PFA which gave a very high autoflourescence
> and 50% ea. MeOH-
> Acetone(ice cold). This method gave me  good (
> already published) pictures
> of ion channels in the t-tubules.
> What I got was confusing results is when I stained
> the cells for actin
> using phalloidin.
> I would greatly appreciate some feedback

=====
Maryse Bailly, Ph.D
Division of Cell Biology, Institute of Ophthalmology
University College London
11-43 Bath Street, London, EC1V 9EL
Tel: (44) 20 7608 4051
Fax: (44) 20 7608 4034
Email: [log in to unmask]

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