Re: [Histonet] Extracellular matrix proteins fixation.

From:Geoff McAuliffe

Hi Diego:

    First off, "well fixed" and a strong immuno reaction are often 
incompatible. The first thing to do is a PubMed search for the exact ECM 
molecules you are interested in. That way you will have peer-reviewed 
material from which to begin work. Or you could try a lot of suggestions 
from the list which may or may not be applicable to your animal, ECM 
molecule, antibody, detection system, etc. Perhaps frozen sections are 
required, or an alcohol-based fixative, etc.
    Good luck!


Diego J. Rodríguez Gil wrote:

>Hi everybody:
>   I am brand new on the list, so I do not know if this question was posted
>before. I apologize for this.
>   I am trying to characterize a family of secreted proteins in the Nervous
>System by immunohistochemistry, and with the regular fixation protocol that we
>used (intracardiac perfusion with 4% PFA), I am not getting any good results.
>Not even after some antigen-retrieval procedures. Before trying random things,
>does anybody have a good protocol to get extracellular proteins well fixed? It
>doesn't matter if it is set up for some other tissue, I would be glad to try
>other protocol(s) if there are any available.
>   Thank you very much for your help,
>   Best regards,
>   Diego.

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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