Standards and Control for PKC IHC

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From:Bruce A Rasmussen <brasmuss@osf1.gmu.edu> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Greetings histoneters,
I am going to attempt to image the distribution of PKC gamma in coronal
rat brain section (20 microns) using immunohistochemistry. The objective
is to get dissociative results between experimental and control animals
with a learning and memory paradigms. Is anyone aware of methods for
creating standards? I am leaning toward the technique of mounting sections
from both control and experimental groups on the same slides and pipetting
the incubating solution directly onto the tissue slices. Any suggestions
on solution volumes and methods to prevent the solution from evaporating?
The protocol calls for incubation with gentle agitation overnight at 4
degrees C. Absolute values for quantification would be gravy but the main
necessity is comparability between batches and antibody economy.
Thanks for your help
Bruce


--------------------------------------
Bruce Rasmussen
Predoc Fellow in Experimental Neuropsychology
The Krasnow Institute for Advanced Study
George Mason University
Mail Stop 2A1
Fairfax, VA 22030-4444
Office:703-993-4358
Lab:703-993-4369
Home:703-765-4570
Fax:703-993-4325
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