Hi Melissa:

    DAKO's rabbit anti-cow GFAP is outstanding. Lasts for years in the refrigerator. I have used it on mouse and rat CNS with beautiful results. On FFPE tissue I use a 1:6000 dilution with a Vector ABC Elite kit but I run the primary Ab overnight and use
nickel+cobalt intensification. On free-floating, fixed frozen sections cut at 20 microns on a sliding microtome the dilution is 1:60,000 (same kit, same intensification, no that dilution is not a typo). Thicker sections, 20-60 microns, are very nice for
astrocytes, you will see much more of the 3 dimensional character of the cells. Paraffin sections that thick are hard to cut, so fix, cryoprotect with sucrose, snap freeze and cut on a cryostat or a sliding microtome. GFAP survives fixation well. The few
times I have done fluorescence I used lower dilutions, about 1:6000 I think, again with Vector reagents. Keep in mind that not all astrocytes are GFAP-positive!
    I have no experience with the anti-NeuN. Go to the PubMed site and look for a recent paper on the subject. Or ask Chemicon, they will have some references. Most of the commercial suppliers keep track of publications with their products, for obvious
reasons.

Melissa Gonzalez wrote:

> Hello,
> Does anyone have any experience using Chemicon's mouse anti-NeuN and/or DAKO's rabbit anti cow GFAP?
> I have seen a few discussions relating to FFPE human tissue, but we are working on mouse frozens for fluorescence microscopy..which leads into my next question, How thick can I keep (or how thin should I cut) the tissue (brain) for optimal visualization?
>
> Thanks in advance for your help!

Geoff
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff@umdnj.edu
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