Re: Acetylcholinesterase

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From:"J. A. Kiernan" <>
To:"Hall, Phil" <>
Date:Wed, 14 Jul 1999 11:45:44 -0400 (EDT)
Content-Type:TEXT/PLAIN; charset=US-ASCII

On Wed, 14 Jul 1999, Hall, Phil wrote:

> It would be nice to be able to use immunochemistry for diagnosis of
> Hirschsprung's disease instead of the enzyme histochemistry method we
> currently use, which needs fresh, frozen sections ...

  Acetylcholinesterase enzyme histochemistry works well on
  frozen sections of formaldehyde-fixed specimens (e.g. fixed
  overnight in formal-calcium or phosphate-buffered formalin, 
  then impregnated with sucrose for cryoprotection). It's
  quite a tough enzyme, and the thiocholine methods are
  extremely reliable. The quantities of hazardous substances
  (I take it you mean anticholinesterases used for inhibited
  controls) used are very much smaller than anything that
  would be toxic to man. The stock needs to be securely stored,
  of course.  

  However, to answer you question, Santa Cruz Biotechnology
  sells three sorts of goat polyclonal anti-AChE of which
  two recognize the human enzyme. US$175 for 0.2 ml.
  They also sell blocking peptides for doing preabsorbed
  controls ($45 a shot). You'd also need an anti-goat secondary 
  detection system, or to use staphylococcal protein A instead
  of the 2ndary antibody.

  The European telephone number for Santa Cruz is their only
  free product:  +800 4573 8000.  They also have a web site: (I suppose that's free too).

 John A. Kiernan,
 Department of Anatomy & Cell Biology,
 The University of Western Ontario,
 LONDON,  Canada  N6A 5C1


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