Re: IHC on hard tissue

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From:Barry Rittman <> (by way of histonet)
To:histonet <>
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		I would suggest that the researcher be convinced that the
sections be
demineralized. While conventional demineralizing agents will interfere with
IHC can use sodium citrate/citric acid or can use Johnnsons PVP method and
many antibodies work with these.
Why is it necessary to retain the mineral? If trying to detect some of the
proteins in enamel then suspect that the hydroxyapatite will not only
prevent adequate penetration of the reagents but may also interfere with
any binding to surface antigens that might occur. If looking at dentin I
think you will, find that many of the antibodies described in the
literature were tested on developing teeth where it was not necessary to
Perhaps an easier task is to educate the researcher, or suggest that such a
technique should be most appropriately tested in a low gravity setting.
NASA is already accepting protocols for the 2004 launches!!

At 07:47 AM 2/18/99 +0000, you wrote:
>Dear all (particularly boneheads),
>One of our researchers wants to:-
>do immunocytochemistry on thin, gound (undecalcified) pieces of
>tooth! (can you believe that?). They will be approx. 150 microns
>thick.  The antibody will be to a naturally occuring protein.
>Any help, experience and (most) suggestions welcomed.  Make that
>Many thanks in advance,
>Diolch yn fawr
>Russ Allison, Wales

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