How does dab staining work

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This kit also includes nickel chloride, which, when added to the substrate working solution, results in a gray-black reaction product. Make a fresh working solution of DAB substrate per instructions.

This indicates the end detection reagents are working appropriately. This kit contains stock solutions in convenient dropper bottles. For additional information, view this product review on Biocompare.

Consideration should be given to tissue type, antigen unmasking protocol, and other detection parameters to achieve the desired staining intensity. DAB brown. DAB-Ni gray-black. AEC red. TMB blue. Vector VIP purple. Alternatively, you can assemble the reagents for DAB staining yourself, using biotin-conjugated secondary antibodies , HRP-polymer-conjugated secondary antibodies , and streptavidin-HRP conjugates.

It is most often used in immunohistochemical IHC staining as a chromogen. It is also used in in situ hybridization ISH and sometimes in dot blots and in western blotting. The oxidized DAB forms a brown precipitate, at the location of the HRP, which can be visualized using light microscopy. The DAB precipitate is insoluble in water, alcohol, and other organic solvents most commonly used in the lab such as xylene and isopropanol.

This allows for great flexibility in the subsequent treatment of the tissue section, such as in the choice of counterstain and mounting medium.

When used together with a nickel or cobalt solution as a DAB enhancer, DAB staining becomes a more intense, black color. These have the following advantages and disadvantages:.

Very simple protocol. Requirement for HRP conjugation of primary antibody. Most heavily used method in research labs. Highly sensitive with low background. Abcam uses this method for the validation of antibodies in IHC.

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Download references. We thank the patients and their families for contributing to this study. You can also search for this author in PubMed Google Scholar. All authors critically edited the manuscript. Correspondence to Charalampos Tzoulis. Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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