Re: asialoglycoprotein antibody

From:Robert Schoonhoven

My experience with many of Santa Cruz's antibodies when used on frozen sections
is that they should be fixed in acetone.

"McTaggart, Ryan" wrote:

> My name is Ryan McTaggart.
> I am attempting to use IHC to I.D. liver cells. hepatocytes, Kuppfer cells,
> cholangiocytes, stellate cells, etc.
> I am starting with Santa Cruz Biotechs (SCB)
> goat asialoglycoprotein antibosy and the
> goat desmin antibody
> my secondary is donkey anti-goat conjugated to Texas red.
> my protocol is not working
> Questions:
> SCB reports using the antibody in fixed tissue with antigen retrieval
> I am using frozen with fixation in methanol shouldn't it work?
> how important is it to use donkey serum in blocking buffer - can i use horse
> - is it the same? I don't need goat serum right?
> how soon do I need to freez the liver after it is removed from the mouse?
> can I let it dehydrate in sucrose for several hours?
> does anyone think antibody staining on a flat, non-agitating surface, with a
> PAP pen is less effective than in a coplin jar with agitation?
> finally, has anyone successfully used these antibodies and are the other
> antibodies you have used successfuly on frozen liver sections to
> specifically I.D. cells?
> thank you for your time - i am a young researcher and your valuable
> experience means a great deal to me.
> best,
> ryan mctaggart
> UCSF - Transplant Research Laboratory

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