[Histonet] problem with DAB staining

[Jan Shivers]

Some suggestions:
1)  your endogenous peroxidase was not quenched properly or entirely
2)  your primary antibody is too concentrated
3)  your secondary antibody is too concentrated
4)  your incubation times are too long
5)  your incubation temperatures are too high
6)  your DAB was mixed up incorrectly
7)  your rinses are insufficient in length
8)  your rinses are insufficient in volume

Jan Shivers
UMN VDL
St. Paul, MN

On Thu, Mar 7, 2013 at 11:50 AM, Nicole Cosenza <ncosenza <@t> siumed.edu> wrote:

> Hello all:
>
> I recently did IHC on paraffin embedded slides via the DAB method. Upon
> addition of the DAB, the entire section turned brown. I know this is not
> true staining, as I don't expect a high yield of my protein of interest.
> Any suggestions as to why this happened and what to do about it?
>
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