Intended use: For In Vitro Diagnostic Use. The BrightDAB substrate DAB is intended for use in immunohistochemistry.
Summary and explanation: The BrightDAB substrate DAB (3,3’Diaminobenzidine) is a widely used chromogen for immunohistochemical staining and immunoblotting. When in the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol and xylene. This product comes in a two-component system consisting of a liquid stable DAB chromogen and DAB substrate buffer. DAB Solution A: Ready–to-Use Buffered H2O2,; DAB Solution B: Concentrated DAB solution. BrightDAB is a very stable and superior formulation of DAB. In some cases, antibodies titers may increase by two-fold. BrightDAB can be used both manually and on automated stainers.The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patient’s clinical history and/or other diagnostic tests. Also available in 500 ml and 1000 ml.
Reagents provided: DAB Solution A: Buffered H2O2 (Ready-to-Use) 110 ml DAB Solution B: Concentrated DAB solution 5 ml
Storage and handling: 2-8°C and in the dark
Principle of method: DAB substrate for use with HRP-labeled detection systems
Reagent preparation: 1 Working solution: Add 40 μl DAB Solution B (± one drop) to 1 ml Solution A, mix well. 2 Incubate the DAB solution one time 8 minutes without washing in between.
Procedure: 1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, polymer HRP, (Labeled polymer; 30 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Substrate (DAB; 8 min), 12. Wash aqua dest (Wash; 2x 2 min), 13. Counterstain, dehydrate and coverslip (Auxiliary)
References: Shan-Rong Shi et al. Applied Immunohistochemistry & Molecular Morphology, vol 7,201-208,1999
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