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A brownish-red precipitate generated by polymerization of the hydrogen peroxide with nitro 3,3'- diaminobenzidine in the treated tissues was observed (Fig.
Diaminobenzidine (DAB); if not available, use leucocrystal violet (LCV) or amido black
The Leica polymer and postpolymer were each incubated for 30 minutes, followed by 10 minutes in diaminobenzidine, and a light hematoxylin counterstain.
12] The slides were then washed, incubated in ABC (Sincere Biotech, Shunyi, China) for 30 min, washed, and then reacted with diaminobenzidine and hydrogen peroxide to visualize the end product.
For double color reaction diaminobenzidine was applied for five minutes and fast red was applied for 20 minutes.
A Horseradish peroxidase-conjugated secondary Ab was used to visualize bound primary Ab and developed using Vectastain ABC kit and diaminobenzidine as a substrate.
Diaminobenzidine tetrahydrochloride was used as a chromogen to localise the peroxidase.
03% diaminobenzidine (Sigma) as chromogen, and the material was counterstained with Mayer's hematoxylin.
The immunoreaction was visualized with diaminobenzidine, and sections were lightly counterstained with Mayer's hematoxylin solution and then dehydrated and mounted.
Processed cells that were reactive were visualized by the presence of a dark brown precipitate when color was developed with diaminobenzidine (DAB).
After being washed with T-PBS, the strips were developed with a diaminobenzidine solution (Sigma).