Re: [Histonet] mouse microglia

From:Geoff McAuliffe

Hi Inga:

   "Mac1" or "F4/80" will stain microglia in frozen sections of  mouse 
brain. Fix by perfusion with buffered paraformaldehyde, remove the brain 
and fix for another 2 hours, no longer (at least for Mac1). Then rinse 
multiple times in buffer, cyroprotect with sucrose, cut 20 micron frozen 
sections, store in PBS, stain free-floating, etc. I have stored sections 
in PBS in the refrigerator for several weeks with no loss of 
antigenicity. Sections air-dired on slides lose microglial antigenicity 
very quickly (2 days). Antibodies for rat microglia will not work on mouse.
    Clint Lincoln has stained mouse microglia in FFPE sections, he used 
a enzyme digestion protocol (pronase?) to unmask the antigenic sites. I 
don't know if I can find his recipe.
    See: Lawson et al., Neuroscience 39:151-170, 1990 and 48:405-415, 
1992 and Perry et al., Neuroscience 15:315-326, 1985.


Inga Hansson wrote:

> Hi all!
> Can anyone suggest a good antibody for microglia that works on frozen 
> and FFPE-tissue in mice?
> Thanks in advance!
> Inga

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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