Cynthia Favara wrote:

> Deb,
>         I know there are antibodies that stain microglia in rats, mouse is a
> different story at least with the strains we study. F4/80 will work fairly
> well on microglia with fresh frozen tissue and I have not difficulty getting
> to stain peripheral macrophages in fixed paraffin embedded tissue.
> Difficulty I have is that because of the network distribution of microglia
> the stains on frozen tissue looks like a huge smear. Vibratome sections may
> be a good alternative but I have not had time to pursue this. Also suppose
> tape transfer would be an option but do not have the instrumentation.
> Mac1 is supposed to work but I have been unable to get specific staining.
>
> Cynthia Favara
> Rocky Mountain Laboratories
> 903 S. 4th Street
> Hamilton, MT 59840
> 406-363-9317
> FAX 406-363-9286
>
> -----Original Message-----
> From: Deb Boswell Lane [mailto:dbl@aretha.jax.org]
> Sent: Friday, November 03, 2000 9:14 AM
> To: Histonet@Pathology.swmed.edu
> Subject: antibody search
>
> We are in search of an antibody against phosphotyrosine (mouse) which
> labels microglia cells and peripheral macrophages.  And an antibody against
> ED2 which labels peripheral macrophages in the rat.  We are looking for a
> mouse equivalent of these 2 antibodies to use on retina.  Has anyone any
> information on these?  Thanks in advance!
>
> Deb Lane
> Deb Boswell Lane
> Professional Biomedical Technologist
> Biological Imaging
> The Jackson Laboratory
> 600 Main Street
> Bar Harbor, Maine 04609
> (207) 288-6193
> dbl@jax.org

    I have had excellent success with Mac1 for microglia in mouse brain.
Fixation by perfusion with 4% paraformaldehyde followed by immersion of brain
slices (5-10 mm) in fix for 2 hours ONLY. Then wash in 3X or more of buffer,
cryoprotect with sucrose and section. Short fixation time is essential!! I use a
Vector Elite kit with either Ni-Co intensification or Vector's own enhancer
(gold chloride I suspect). Dilution of the primary Ab is 1:2000 overnight at
room temp.
    Contact me if you need more information.

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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