2. DAB Chromogenic Kit, 200 T, $150

Product Information

Description

DAB chromogenic kit, which will be obvious color by horseradish peroxidase (HRP), is used in experiments such as immunohistochemistry, in situ hybridization, and Western blotting (WB). DAB is a substrate for HRP, and produces a brown precipitate catalyzed by HRP, which is insoluble in water and ethanol. Therefore, subsequent dyeing with alcohol-soluble dyes can also be performed after DAB chromogenic reaction. In immunohistochemistry or situ hybridization experiments, this kit can detect a total of at least 200 samples using 50 μL of chromogenic solution for each sample.

Storage and Handling Conditions

Transport in wet ice; store at 4°C away from light, valid for 12 months.

Component

Assay Protocol

1. Preparation of DAB working solution: 20 μL of 50×DAB stock solution is added to each 1 mL of DAB diluent solution. The resulting solution should be mixed well for use, which needs to be prepared and used immediately and be limited to use on the same day. It is not recommended to store it for a long time.

2. The DAB working solution is dropped directly on the processed immunohistochemical sections or WB membranes. Then the sections or membranes are incubated at room temperature in the dark for 3-30 min or longer until the desired color develops.

3. DAB working solution on sections and membranes should be removed, and the chromogenic reaction would be stopped by washing twice with PBS or distilled water.

Note

1. If the background is too dark during immunohistochemistry, the following measures could be considered: prolonging the washing time, using an appropriate blocking solution for blocking, inactivating endogenous catalase, shorting the color development time, and reducing the concentration of secondary antibodies.

2. If there is no chromogenic reaction or the color is too light, the following measures could be considered: increasing the concentration of the primary antibody and the secondary antibody appropriately, prolonging the chromogenic reaction time, and checking whether the secondary antibody is normally chromogenic.

3. Please be careful when handling as DAB is harmful to the human body. It is necessary to take precautions to prevent DAB from coming into direct contact with the human body or being inhaled.

4. It is recommended to wear a lab coat and disposable gloves when operating.

For Research Use Only!