Re: histone immuno

From:"J. A. Kiernan"

I missed the original question, so am commenting on 
the basis of a reply (cited below).  My comment may 
therefore be irrelevant or inappropriate!

If Ronald wants to stain _all_ histones, he doesn't need
to use expensive antibodies and associated immunostuff.

An anionic dye at pH 8.5 or higher is quite selective
for strongly basic proteins. Apart from histones the only 
other ones I can think of in animal tissues are some
cytoplasmic secretory granules and stuff in sperm tails.
Biebrich scarlet is a good dye for this sort of job
because its colour stays the same over a wide pH range.

The staining of histones in cell nuclei is enhanced if
you first extract the DNA by treating the sections with
DNase (won't work after formaldehyde; fine after Carnoy)
or with trichloroacetic acid (nasty stuff; works after
formaldehyde, but expect section losses).

These olde worlde methods for nucleohistones are described
and critically discussed in histochemistry textbooks,
especially the older ones such as Lillie. I've done some
of them myself, and can testify that they work. The only
difficulty is keeping sections on slides. Extra-strong
chrome-gelatin was better than some other adhesives, and
better, surprisingly, than APES-treated slides.

I'll be happy to provide references to anyone interested
in non-immuno staining of histones. 

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

Tamara Howard wrote:
> Ronald -
> There are quite a few histone antibodies available commercially, but I
> don't know if there is a general "anti-histone" one. The Abs I've used are
> histone-specific. You can also get fancy and buy phosphorylation state-
> and site-specific antibodies. Upstate Biotechnology is one vendor for the
> phospho-specific antibodies.
> Tamara
> |--------------------------------------------------|
>  Tamara Howard
>  Department of Cell Biology and Physiology
>  University of New Mexico - Health Sciences Center
>  Albuquerque, NM 87131
> |--------------------------------------------------|

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