RE: Microglial cell marker

From: McCoy, J. Philip (NIH/NHLBI) (McCoyJ@nhlbi.nih.gov)
Date: Mon Nov 25 2002 - 09:16:10 EST


If you do flow cytometry with lectins, beware of the problems caused by
agglutination. For the GS-IB4, you are much better off using the GS-IA3B1
lectin, and blocking the activity of the A subunit with appropriate sugar.
This yields a functionally monovalent probe and avoid agglutination of
cells. For details, see:

McCoy JP, Shibuya N, Riedy MC, and Goldstein IJ: Griffonia simplicifolia I
Isolectin as a Functionally
	Monovalent Probe for Use in Flow Cytometry. Cytometry 7:142-146,
1986.


-Phil

J. Philip McCoy, Jr., Ph.D.
Director of the Flow Cytometry Core Facility
NHLBI - NIH
Bldg 10, Rm 4A07
10 Center Dr., MSC 1357
Bethesda, MD 20892


> -----Original Message-----
> From: Richard Haugland [mailto:richard.haugland@probes.com]
> Sent: Thursday, November 21, 2002 7:54 PM
> To: Cytometry Mailing List
> Subject: Re: Microglial cell marker
>
>
> Dr. Wes Chang of our place provided me this potential answer.
>
>
> GS-IB4 (Grifonia simplicifolia) and RCAI and RCA120 (from
> Ricinus communis) are
> two lectins that are reported to stain microglia in a variety
> of species,
> although I do not know about cat specifically. For
> antibodies, I found a
> message on the Histonet server that indicates DAKO's CR3/43 antibody
> labels microglia in human brain tissue. Again, don't know about cats.
>
> We, and others, make fluorescent GS-IB4. (If the image
> reproduces the green
> fluorescence is the microglia of rat brain, the red fluorescence Nissl
> substance and the blue fluorescence DAPI in nuclei.)
>
>
> [Image]
>
>
>
> "Richard K. Meister" wrote:
>
> > Hello, everyone:
> >
> > I have a colleague who wants to study microglial cells in
> cat brain.  Do
> > any of you know of a marker for microglial cells?  (I've
> looked, but so far
> > have only found MHC II markers used to study activated
> microglia.)  We
> > would prefer an antibody that is specific for cat or
> cross-reacts, but any
> > other that we could test for cross-reactivity would also be helpful.
> >
> > The application would be to stain either parrafin-embedded
> tissue or frozen
> > sections.  I would also like to hear about any markers that
> would work for
> > flow cytometry.
> >
> > Thanks in advance,
> >
> > Rick Meister
> >
> > *  *  *  *	*  *  *  *  *  *  *  *	*  *  *  *  *  *  *  *
> *  *	*  *  *
> > *  *  *  *	*  *  *  *	 *
> > *  Richard K.
> > Meister		     Email:  meister.1@osu.edu		      *
> > *  The Ohio State University	   Voice:  (614)
> > 292-9716		       *
> > *  Dept. of Veterinary Biosciences	   FAX:    (614)
> 292-6473	     *
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