Re: Subject: versican, decorin, and dendritic cells
|From:||"Brothers, Rachel" <firstname.lastname@example.org>|
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.
Rachel Stock HTL(ASCP)
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
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.
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