The DAB Chromogenic Reagent Kit is designed for detecting targets in immunohistochemistry (IHC), in situ hybridization, and membrane-based assays such as Western, Southern, Northern blotting, and EMSA, utilizing Horseradish Peroxidase (HRP).
In this reaction, 3,3'-Diaminobenzidine (DAB) is oxidized by HRP in the presence of a peroxide substrate, generating an insoluble brown precipitate at the sites where the HRP-conjugated antibody is bound. This precipitate is insoluble in water and ethanol, making it compatible with standard dehydration and mounting procedures for permanent preservation.
Kit Contents
D1505853 | Components | 20ml | 100ml | Storage |
D1505853A | DAB Chromogenic Solution A | 10ml | 50ml | -20℃. Store in the dark. |
D1505853B | DAB Chromogenic Solution B | 10ml | 50ml | -20℃. Store in the dark. |
Usage method
1. For tissue sections or protein blotting membranes, after incubation with horseradish peroxidase (HRP)-labeled antibodies or other forms of probes, wash them 3-5 times with the appropriate washing solution for 3-5 minutes each time.
2. Mix solution A and B in a 1:1 ratio to obtain the DAB working solution. It is recommended to use it within 4 hours.
3. Add an appropriate amount of DAB staining working solution to the tissue sections or membranes to ensure full coverage of the samples.
4. Incubate at room temperature in the dark for 3-30 minutes or longer (up to 24 hours) until the color development reaches the desired depth.
5. Remove the DAB staining working solution and wash 1-2 times with distilled water to terminate the staining reaction.
6. For tissue sections or cell samples, after the staining reaction is terminated, they can be re-stained with other dyes (commonly hematoxylin staining solution). For membranes, after the staining reaction is terminated, they can be air-dried at room temperature and stored in the dark.
Note：After thawing, the solution can be aliquoted as needed for the experiment. If the solution turns brown, it indicates that the active ingredient of this component has deteriorated and should not be used.
Matters needing attention:
1. DAB is harmful to the human body. Please handle it with care and take effective protective measures to avoid direct contact with the body or inhalation into the body.
2. This product is for scientific research only. Do not use it for medical purposes, clinical diagnosis or treatment, food or cosmetic purposes. Do not store it in ordinary residential areas.
3. For your safety and health, please wear laboratory coats and wear disposable gloves and masks when operating.
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