DAB Peroxidase Substrate Kit (Purple-Blue Color) 

3, 3’-Diaminodbenzidine (DAB) is a chromogenic substrate that can be used with horseradish peroxidase (HRP) for immunohistochemistry and immunoblotting applications. Under HRP catalysis, the substrate DAB reacts with hydrogen peroxide, transferring electrons from DAB to the peroxide, producing an insoluble brown product.

MCE DAB Peroxidase Substrate Kit (Purple-Blue Color) has an upgraded formula based on the original, enhancing the sensitivity of color development. The final color will appear gray-blue or purple-blue. This kit can be used for staining and color development detection in experiments such as immunohistochemistry of cells or tissues, in situ hybridization, Western blotting, and for visualizing endogenous HRP in cells or tissues.

-20℃, 1 year

Keep away from light and avoid repeat freeze-thaw cycles.

1. After incubation: Incubate regular tissue sections, cell samples, or membranes with HRP-labeled antibodies or other types of probes, and then wash 3-5 times with the appropriate washing buffer. For tissues or cell samples detecting endogenous HRP, wash them 3-5 times with washing buffer after appropriate fixation, each time for 3-5 min.

2. Prepare DAB working solution: Refer to the table below for preparation (adjust volume as needed).

Note: DAB Concentrate (20×) and DAB Diluent Buffer must be thoroughly mixed to prepare the working color development solution.

3. Incubation with DAB: After washing, remove the washing solution and add the DAB working solution to fully cover the sample. Incubate at room temperature in the dark for 1-30 min or longer.

Note: 1) If no background appears, incubation can continue until the desired color intensity is reached.

2) In Western blot experiments, ensure there is enough DAB working solution for the membrane to move freely.

4. Stop the reaction: Remove the DAB working solution and wash several times with ddH₂O to stop the color development reaction.

5. Post-staining and mounting: For cells or tissue samples observed under a microscope, optional counterstaining can be performed, followed by mounting in an aqueous medium. Alternatively, samples can be dehydrated and mounted in an organic mounting medium. For Western blot experiments, wash the membrane with ddH₂O, air dry, and store at room temperature.