Out laboratory used Dako´s AEC substrate up until two months ago.

Since it wasn´t available in our region, we instead received the DAB products.

In all cryo liver staining attemps however, we´ve encountered difficulties.

The results are inhomogenous, the background highly overstained and the signal either too strong or not detectable.

Even an incubation time of 1 minute with the substate solution is enough to cause this background oversatiation.

We´ve tried to modulate factors such as antibody dilution, incubation time with the substrate, the kind of primary antibody used, substrate diluation but for weeks the results have been the same.

Is there anyone with similar problems?

If so, how did you solve them?

Is there maybe a rule of thumb we are not aware of when to preferably use AEC/ DAB products?