RE: help - IHC

From:"Johnson, Teri"


I've found that yes, SC antibodies are optimized for frozen section.  Sometimes they work in paraffin, sometimes they don't.  Their new catalog has a listing of the ones which have been tested in paraffin embedded material, and you might want to try using a different catalog number if they offer it.  On the integrins, you might try switching to a polyclonal, although it looks like the catalog doesn't offer any optimized for paraffin-embedded sections.

You could try different digestion techniques, including proteases and HIER with different solutions.  I'd be tempted, though, to try it on cryostat sections first to verify it stains properly before doing anything different.

Also, if possible, try using a different fixative (zinc formalin or glyoxal) to see if your specimen retains antigenicity if the frozen section staining shows promise.

Sorry, but it's the best advice I can give.  Hope it helps!

Teri Johnson
Managing Director Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110

-----Original Message-----
From: Mcleod, H, Heather [mailto:hmcleod@CHEMPATH.UCT.AC.ZA]
Sent: Wednesday, April 17, 2002 11:41 AM
Subject: help - IHC

Dear Histonetters
I am trying, without much success to stain ffpe sections with the 
following antibodies
(1) Integrin alpha-6 (BQ16) Santa Cruz sc-13542 Mouse monoclonal and 
(2) Fibulin-1 (C-16) Santa Cruz sc-8675 Goat polyclonal
I'll be grateful for advice from anyone who has used these ab's on 
ffpe material.
I am told that all SC antibodies are optimised for / on  frozen 
sections.  What is the trick to getting such antibodies to stain 
fixed material?  
Heather McLeod (GSH / UCT)

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