1) 5mL concentrated amber-colored DAB Chromogen Solution.
2) 200mL clear Stable DAB/Plus Substrate Buffer.
3) One empty mixing dropper bottle.
Aliquot 1mL of Stable DAB/Plus Buffer in mixing bottle. Add 20μL (one drop) of
concentrated Stable DAB/Plus Chromogen. Replace tip and mix.
1.) After peroxidase incubation, wash tissue sections with wash buffer.
2.) Wipe slides removing excess buffer. Add enough drops of working Stable DAB/Plus
solution to cover tissue sections.
3.) Incubate for 5-10 minutes at room temperature.
For optimal results, observe reaction under the microscope for signal development.
Once the desired signal to noise ratio is achieved, stop the reaction by washing slides in buffer.
Note: The working Stable DAB/Plus solution is stable for at least 2 weeks and should be
prepared in an opaque bottle. Store at 2-8ºC when not in use. Any solution not used after
this period should be discarded.