Re: Permeabilization with ethanol

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From:Geoff McAuliffe <mcauliff@UMDNJ.EDU> (by way of histonet)
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Paul Klosen wrote:

> A 16:45 08/11/1999 EST, a écrit :
> >Histonetters,
> >I am looking for a reference for using buffered, graded ethanols for
> membrane permeabilization that is a little less destructive than saponin or
> other
> >methods. A detailed method would suffice or the exact reference you use.
> Any and all replies will be appreciated.
> >Thanks in advance,
> >Nancy Lemke
> Hi,
> I have used buffered ethanol membrane permeabilization for ICC with very
> good results, at least in terms of permeabilization. I don't know whether
> it is less destructive than saponin. Ethanol being a precipitating
> fixative, ethanol permeabilization may indeed precipitate some membrane
> proteins (or even cytoplasmic proteins) that would be extracted by the use
> of detergents as saponin.
> We use a procedure derived from the following paper:
> Eldred WD et al. (1983) J Histochem Cytochem 31 (2): 285-292
> I have never looked at the ultrastructure of the tissue, which according to
> these authors should be quite OK.

    I have looked at ultrastructure in the CNS and found that different regions
responded differently, some looked quite nice, some rather chewed up.
    Just my $0.02 worth.

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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