What everyone else is saying is true and you will need to check your
procedure carefully.  Manual staining is a technique that is difficult
with some antibodies and especially when working with a large slide
number.

One of the things that I did was eliminate using a pap pen.  I used 2
X 2 strips of parafilm that I cut in strips of 1 X 2 and layed them on
the tissue/slide after the reagent has been applied.  To remove I
dipped the slides in buffer and the film would slide right off.

Good luck.  I did IHC manually for a long time and it is not always easy.

Vikki

On 5/9/08, Karla Arrington  wrote:
> I do Immuno's by hand and I am getting background DAB precipitate on the inside circle of my sections.
> What is causing this and how do I get rid of it?
>
>
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