I used to do this in skin sheets, I have also stained isolated hair
follicles. I think your samples should fit into 6 well plates. I used to
incubate overnight at 4C for the primary antibody, otherwise the method is
very similar to that for sections. Remember to use plenty of each reagent so
that your samples remain immersed at all times. You can transfer the
reagents with pasteur pipettes or if your tissue is robust enough transfer
it to a new well. When I stained hair follicles, I used a 3:1 solution of
ethanol:acetic acid to permeabilise the samples.

I would optimise the staining on frozen sections prior to starting on the
whole mounts. I have no experience of staining whole embryos, so I'm not
sure how well your reagents will penetrate the samples. No doubt someone
else will give you an answer.

Good luck

Margaret

-----Original Message-----
From: Hernan Aldana Marcos [mailto:haldana@unimoron.edu.ar]
Sent: Wednesday, March 16, 2005 8:28 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] immuno in block


Dears
I need to make immunos in the whole retinas. Can anybody have a protocol to
do it?

Or a similar technique in whole embryos. Times, concentrations or any
advices to make Immunohistochemistry in  thick tissues.

Thanks in advance

 


Dr. Hernán J. Aldana Marcos
Facultad de Medicina. Universidad de Morón
Machado 914. B1708JPD. Buenos Aires. Argentina 
e-mail alternativo hernanjavier@yahoo.com
web: http://hjaldanamarcos.bravepages.com
http://histologia.bigthicketdirectory.net/main.html

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