Oligodendrocyte staining

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From:Kelly Randall <kellyr@neuro.hfh.edu> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Hi there everyone!

I have been trying to stain oligodendrocytes
with 2 separate antibodies (that are supposed to be the easiest and most
reproducible).

On our fixed, paraffin embedded rat brain tissue (6um thick sections).
CNPase (from Sternberger Monoclonals) - This stains oligos and myelin -
have good staining using DAB substrate, but it will be very hard to perform
image analysis due to the intensity of the myelin staining.

On frozen rat brain tissue (20um thick sections)
APC  (from Oncongene- ithink).  I have been using Cy3 to localize, and
have had only mediocre results.  We are hoping to get this antibody
working in paraffin- but so far have not had any decent staining.

I still have some "tweaking" to do but would like to know if any one out
there in Histoland has been using anything else to detect oligodendrocytes.
I have read of a number of other antibodies (i.e. galactocerebroside,
tau, etc) but from what i have heard these are all very hard to use.

Thanks in advance for any tips or suggestions!

Kelly Randall




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