I am performing IHC with the ABC elite kits using Impact DAB, both from Vectorlabs. My problem is that the DAB solution precipitates when added to my intestine samples on the slide, like tinsel on a xmas tree. They are in solution but they happen to attach to my tissue. The first picture is the precipitate not over tissue, the second is the precipitate over the tissue, and the third is a tissue section that did not have any precipitate over it, so while I consistently see a precipitate, it does not cover the tissue and slide uniformly. Any suggestions to eliminate the problem.
Hi Marcus, I work with ABC elite kit and DAB (sigma) on brain slices, we almost have no problem of DAB precipitation. It is interesting though that both precipitates not over tissue and over tissue in your cases are quite similar. So I suspect the problem might come from the reaction solution or the slides. Good luck.
Hi Marcus,
DAB solution is a very unstable solution and can react pretty quickly. I suspect that your solution is gone off. You should check it for precipitates by simply putting a droplet on a glass slide. If you see any particles precipitating without any tissue and reaction than that means you will need to buy a new kit. Alternatively you may try to filter it but this may not give you reliable or consustent results.
Best wishes,
Refik
To all who probably knew the answer anyways, it turns out that the mouse gut intestine is sticky for the ABC reagent, so I needed to add additional washing steps to alleviate this problem. Thank you to Vectorlabs for trouble shooting this for me.
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