Hi Lorrane,
Not sure if this helps, but the following are some replies I received 
regarding rat macrophages markers (a couple years ago now).
Cheers! Greg

Date sent:      	Fri, 02 Oct 1998 09:18:07 -0400
From:           	"Hewlett Bryan  (CMH)" 
<HEWLETT@EXCHANGE1.CMH.ON.CA>
Subject:        	RE: Rat Macrophage - Staining Problems
Forwarded to:   	DOBBIN@acad1.cs.upei.ca
To:             	"'Histonet@Pathology.swmed.edu'" 
<Histonet@Pathology.swmed.edu>,
	'Dave Tacha' <dtacha@ncal.verio.com>

Dave,

10-20 micrograms/ml sounds a little high to me, we seldom go 
above 5 and
with HIER our range is usually 0.1- 2.0 micrograms/ml.
Am not familiar with the antibody you use.

We use mouse anti-rat monocyte/macrophage clone ED-1 from 
Serotec
cat#MCA341. For formaldehyde fixed paraffin sections of rat lung, 
we use
HIER in 0.01M citrate buffer pH6.0 for 10 min.@ 96C. ED-1 is 
diluted
1:250 in 1% v/v normal swine serum in 0.05M TBS pH7.6, 
incubation is 1hr
@RT, detection is with biotinylated rabbit anti-mouse( absorbed 
against
rat) (Dako code E0464, diluted 1:300 in 1%v/v NSS/TBS, 
1hr@RT), followed
by Streptavidin/Pox(Dako code P0397, diluted 1:600 in NSS/TBS, 
1hr@RT).
Chromogen is AEC, if you use DAB you may find that the primary 
can be
further diluted to 1:500 and if you use intensified DAB then 1:1000! 
Regards

Bryan



>----------
>From: 	Dave Tacha[SMTP:dtacha@ncal.verio.com]
>Sent: 	October 1, 1998 8:13 PM
>To: 	Histonet@Pathology.swmed.edu
>Subject: 	Rat Macrophage - Staining Problems
>
>We been using a "mouse anti-rat macrophage" from Pharmagen.  
We
>incubated the rat tissue (lung) for 1 hour at 10 and 20 micrograms 
per
>ml.  I also pretreated the tissue with and without antigen retrieval.  I
>used Pharmagen's goat anti-mouse with minimal cross-reactivity 
to rat.
>However, we are not getting any staining.
>
>Could someone help me with a procedure, or technical hint; or 
another
>perhaps another rat macrophage antibody that works in formalin-
fixed
>paraffin embedded tissues?
>
>My main concern is that the secondary antibody may not 
recognize the
>primary antibody.
>
>Has anyone ever heard of MOMA?
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Date sent:      	Tue, 29 Sep 1998 21:52:40 +0100 (BST)
From:           	Richard Edwards <REE3@leicester.ac.uk>
Subject:        	Forwarded: Re: ED1 antibody
Forwarded to:   	DOBBIN@acad1.cs.upei.ca
To:             	dobbin@upei.ca
Priority:       	normal

   Dear karen,
     
     I have used ED1 and ED2 to stain macrophages in rat 
tissues(spleen,
     lymph node and liver...). If you want to stain Kupffer cells in
liver, 
     ED2 will stain very well. I didn't try ED1 in liver but I assume it
     works as well as ED2. In general, ED1 expression increses with 
     activation and ED2 expressed by mature resident 
macrophages. 
     
     I purchased both antibodies from Biosource(800-242-0607) in 
ascites
     form. It works very well.
     
     I hope this information helps!
     
     Lina Li
     lil@pfizer.com


______________________________ Reply Separator 
_________________________________
Subject: ED1 antibody
Author:  HistoNet@Pathology.swmed.edu at Internet
Date:    9/16/98 7:36 AM


Hi histonetters
     
Does anybody know of an antibody to ED1--it staines a 
cytoplasmic antigen
on 
the majority of mononuclear phagocytes including monocytes .  
There is 
another 
antibody KU-1 that recognizes a cytoplasmic antigen on tissue
macrophages.
     
I thought perhaps either of these might stain Kupfer cells in rat liver.
     
Thanks to you all in advance.
     
Karen WIghtman
Eli LIlly & Co
Greenfield, IN 46140
ph 317-277-4136
FAX 317-277-4954
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From:           	"Greg Dobbin" <AVCN1/DOBBIN>
Organization:   	University of P.E.I.
To:             	dobbin
Date sent:      	Tue, 29 Sep 1998 08:02:59 -0400
Subject:        	(Fwd) Re: Seeking Ab to Rat Macrophages


Greg 
            The  anti-lysozyme  antibody  from  Dako works   on  rat 
alveolar  
macrophages.............also  their  anti-alpha-1-
antitrypsin.................but not  
so good..................in  terms  of the  numbers  of  positive 
macrophages.
                                  Cheers
                                            Richard  
Edwards..................... 
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
 Date sent:      	Mon, 28 Sep 1998 11:48:16 -0700
From:           	Patsy Ruegg <rueggp@Path1.UCHSC.edu>
Subject:        	Re: Seeking Ab to Rat Macrophages
Forwarded to:   	DOBBIN@acad1.cs.upei.ca
To:             	Greg Dobbin <dobbin@upei.ca>
Organization:   	UCHSC - Dept. of Pathology
Priority:       	normal

Pharm
in is your best bet
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Date sent:      	Mon, 28 Sep 1998 11:19:20 -0600
From:           	Gayle Callis <uvsgc@msu.oscs.montana.edu>
Subject:        	rat macrophage antibodies
Forwarded to:   	DOBBIN@acad1.cs.upei.ca
To:             	histonet@pathology.swmed.edu

Contact SeroTek, they have an excellent selection of rat 
macrophage
subsets, ED1, ED2, ED3.  

Gayle Callis
    
     
     


>
>
>
>
>
>
Greg Dobbin
Pathology Lab
Atlantic Veterinary College, U.P.E.I.
550 Unviversity Ave.
Charlottetown, P.E.I.
Canada,  C1A 4P3
Phone: (902)566-0744
Fax: (902)566-0851