Re: Subject: versican, decorin, and dendritic cells

From:"Brothers, Rachel" <rachel_stock@internet.organo.com>

Mark, 
I have worked out some proteoglycan IHC's and versican and decorin. 
The protocols for both decorin and versican use the Vector Elite kits. 
    1. Antigen Retrieval Solution: (2X) 4.2g Citrate in 500ml dH2O + 500ml 0.1M
NaOH, pH 6.0
    For final mix: 1:1 with dH2O on day of experiment. Place slides face-up in a
small plastic box, and cover with 1 inch of Antigen Retrieval Solution.  Place
the box in 1 inch deep H2O in a glass tray. Microwave on high for 4 minutes (no
boiling!). Cool 1-5 minutes, then rinse. 
    2. Incubate in endogenous peroxidase blocker (Peroxoblock) and rinse
3. Serum block from kit,  incubate for 20-45 minutes at room temperature. Rinse.
    4. Try a primary dilution of 1:250, 1 hour incubation at room temperature (I
used primaries made by Richard Gallo, formerly of Boston's Children's Hospital,
but these antibodies are available commercially.) 
5. Proceed as directed by the Vectastain kit. 
6. I use DAB with a Nuclear Fast Red counterstain

As for dendritic cells, the only one I use is a CD1a stain for Langerhans cells
in skin. I can send you a protocol if you'd like. 
Best regards, 
Rachel Stock HTL(ASCP)
Organogenesis, Inc.
150 Dan Road, 
Canton, MA 02021

Ph. (781) 575-0775

Date: 2 Nov 2000 19:25:21 -0600
From: "Mark Elliott" <MElliott@mrl.ubc.ca>
Subject: antibodies-biglycan, versican, decorin, and dendritic cells

Hi
Was wondering if anyone knows of a commercial source for 
antibodies to biglycan, versican and decorin that would work on 
Formalin-fixed, paraffin-embedded human tissues.
In addition, we are trying to localize dendritic cells in FFPE human 
lung and were wondering if anyone could receommend a good 
antibody which is specific for these cells.
Thanks for your help.

Mark




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