Re: Lost DAB replies
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
> Would you know why does the DAB solution precipitate when applied on the
> tissue. I have been always using Vector DAB kit. This usually happens when
> the staining is done, so it doesn't interfere with the results. I wash my
> sections with PBS before applying the DAB solution.
If you dissolve DAB directly in buffer (pH 7.2-7.6), you get
an opalescent solution because the sparingly soluble free base
is released and is reluctant to redissolve. To avoid this,
dissolve the DAB hydrochloride first in a small amount
(about 1 ml) of distilled water, and then mix this solution
with the larger (50 ml or whatever) volume of the buffer.
If you do it that way there is never a precipitate.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
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