Re: limits of torture (to tissues in IHC)
No problem to do triple IHC with BrdU as one of the antibodies. I usually
treat with HCL 0.2N, 15', 37deg, then wash 15 min with borate buffer, and
then PBSx3. Most of the antibodies do not mind all this treatment, so I
block/permeabilize and then incubate with a mixture of the BrdU antibody
and the other two antibodies as usuall. Just remember that nuclear proteins
need some extra permeabilization, for example 0.3-0.4% Tx100; BSA 3%, NGS
5%, 30' in 37deg and then 2hrs or ON. Also I found Texas Red (1:800) to
work really well with the anti BrdU antibody from Accurate.
At 09:16 AM 7/25/2001 , you wrote:
> This to do with double or triple IHC labelling
>studies. For eg If one has to detect BrdU together
>with a cytoplasmic protein. BrdU incorporated in the
>DNA will be visible only after some DNA denaturation
>(1-4NHCl, or DNAase or pepsin etc). Wouldn't this
>destroy the structure of the protein that one is
>trying to detect. Is there any solution.
>Sunil Thomas K
>Do You Yahoo!?
>Make international calls for as low as $.04/minute with Yah
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