Re: antibody/Mac1

From:Geoff McAuliffe <mcauliff@UMDNJ.EDU>

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 []
> Sent: Friday, November 03, 2000 9:14 AM
> To:
> 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

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

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