Re: limits of torture (to tissues in IHC)

From:Gilmor Keshet

Hi Sunhil,
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

<< Previous Message | Next Message >>