RE: Specific markers for macrophages in culture

From:Greg Dobbin <dobbin@Upei.CA>

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)" 
Subject:        	RE: Rat Macrophage - Staining Problems
Forwarded to:
To:             	"''" 
	'Dave Tacha' <>


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 
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 
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 
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, 
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! 


>From: 	Dave Tacha[]
>Sent: 	October 1, 1998 8:13 PM
>Subject: 	Rat Macrophage - Staining Problems
>We been using a "mouse anti-rat macrophage" from Pharmagen.  
>incubated the rat tissue (lung) for 1 hour at 10 and 20 micrograms 
>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 
>perhaps another rat macrophage antibody that works in formalin-
>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 <>
Subject:        	Forwarded: Re: ED1 antibody
Forwarded to:
Priority:       	normal

   Dear karen,
     I have used ED1 and ED2 to stain macrophages in rat 
     lymph node and liver...). If you want to stain Kupffer cells in
     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 
     I purchased both antibodies from Biosource(800-242-0607) in 
     form. It works very well.
     I hope this information helps!
     Lina Li

______________________________ Reply Separator 
Subject: ED1 antibody
Author: at Internet
Date:    9/16/98 7:36 AM

Hi histonetters
Does anybody know of an antibody to ED1--it staines a 
cytoplasmic antigen
the majority of mononuclear phagocytes including monocytes .  
There is 
antibody KU-1 that recognizes a cytoplasmic antigen on tissue
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

            The  anti-lysozyme  antibody  from  Dako works   on  rat 
macrophages.............also  their  anti-alpha-1-
antitrypsin.................but not  
so  terms  of the  numbers  of  positive 
 Date sent:      	Mon, 28 Sep 1998 11:48:16 -0700
From:           	Patsy Ruegg <>
Subject:        	Re: Seeking Ab to Rat Macrophages
Forwarded to:
To:             	Greg Dobbin <>
Organization:   	UCHSC - Dept. of Pathology
Priority:       	normal

in is your best bet
Date sent:      	Mon, 28 Sep 1998 11:19:20 -0600
From:           	Gayle Callis <>
Subject:        	rat macrophage antibodies
Forwarded to:

Contact SeroTek, they have an excellent selection of rat 
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

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