[Histonet] Post immunofluorescence immunostaining

Gayle Callis gcallis <@t> montana.edu
Wed May 26 09:23:03 CDT 2004

I suppose one could pull the coverslip at risk of damaging section and then
come back with antiPE, antiFITC, or go to Molecular Probes and see if they
have an antiAlexa 488 antibody. There are some nice antibodies for
detecting fluorophores out there.  

I would suggest coverslipping with PBS for viewing with fluorescent
microscope and do this immediately, without using any coverslip edge
sealant.  Soak off coverslip and try immunostaining.  DAKO has an excellent
Rabbit antiFITC, (do not use the rabbit antiFITC-HRP conjugate, it doesn't
work as well per Chris van der Loos suggestion/experience) then come back
with an antiRabbit secondary (Donkey antirabbit-biotin) etc. Just make sure
you are careful with blocking and secondary hosts to prevent nonspecific
cross reaction detection.

At 03:05 PM 5/26/2004 +0200, you wrote:
>When performing immunofluorescence can you then add a substrate or any thing
>else to turn this into a non fluorosecent slide which can then be visible
>down a light microscope.
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
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