On 9 Aug 2002 17:01:23 -0500
Carol Bobrowitz  wrote

Does anyone use Fibronectin or Collagen III on paraffin sections?
What pretreatments do you use?  (Heat pH 6.0 or 10.0 or enzyme digestion.)
Thank you,

Carol, in my laboratory I use the DAKO Polyclonal Fibronectin antibody cat
no A0245. Pretreatment is HIER using citrate pH6.  Dilution of 1 in 400.
Incubation time of 1 hour with a LSAB detection system. Control tissue is
normal kidney where the fibronectin will stain the tubular basement
membranes.

The collagen type III antibody that I use is a Biogenix Monoclonal cat no
MU167UC. Pretreatment is HIER using Tris-EDTA pH8. Dilution is 1 in 100.
Incubation time of 1 hour with a LSAB detection system. I worked this up
using normal liver as a control. If your doing liver work the EDTA retrieval
can give some endogenous biotin issues which may require blocking. 

Hope this helps,

Regards

Mark Donovan
Anatomical Pathology
The Alfred Hospital
Melbourne, Australia.




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