The immunoreaction was amplified with the avidin-biotin complex and visualized with 3,3'-diaminobenzidine
For the most part, the vast majority of microscopic pathology is still done by viewing or imaging one marker at a time in serial sections, using a hematoxylin counterstain and either eosin (H&E) or, for protein IF1C measurements, 3,3'-diaminobenzidine
(DAB) as a chromogen.
Bound antibodies were detected with goat anti-rabbit IgG labeled with peroxidase diluted to 1/500 in PBS and 3,3'-diaminobenzidine
Specific primary antibody binding was detected using EnVision+ (Dako North America, Carpinteria, California) with 3,3'-diaminobenzidine
tetrahydrochloride as chromogen.
Following several additional washes, sections were treated for 10 minutes with freshly prepared reaction mixture, containing a 10 mg tablet of the substrate 3,3'-diaminobenzidine
tetrachloride (DAB, Sigma, St.
We used this molecule as a model to establish optimal conditions for detecting RCPs with HRP and 3,3'-diaminobenzidine
tetrahydrochloride (DAB) staining.
Slides were again washed and incubated with 3,3'-diaminobenzidine
(DAB) chromogen, washed with [H.
The reaction was revealed by 3,3'-diaminobenzidine
and counterstained with hematoxylin.
After washing in PBS, the sections were treated with a mixture of 3,3'-diaminobenzidine
(DAB; Sigma, St.
E-HRP binding was visualized by the precipitate formed when reacting with 3,3'-diaminobenzidine
tetrahydrochloride (DAB) in the presence of urea hydrogen peroxide and Co[Cl.
In a positive reaction, the surface of the fish egg was stained brown as a result of the oxidation of 3,3'-diaminobenzidine
(substrate) by horseradish peroxidase bound to the egg surface by the antibody.
Staining was then visualized by using 3,3'-diaminobenzidine