Diaminobenzidine (DAB) is oxidized by horseradish peroxidase (HRP) to give a brown color. It is a sensitive colorimetric substrate used with HRP in immunohistochemistry (IHC) and immunoblotting applications. This DAB Substrate Kit has improved stability, ease of formulation, and greater sensitivity.
- DAB stock solution (50×)————-3 ml
- Stable H2O2 solution (50x)———–3 ml
- DAB buffer——————————–120 ml
Store at 2-8oC.
Recommended Assay Procedure for Immunohistochemistry
- Before DAB staining, add 40 μl of DAB stock solution and 40 μl of stable H2O2 solution for every 2 ml of DAB buffer. The mixed DAB substrate solution can be stored for up to 6 hours. Storing the reagent for longer periods of time will cause precipitate formation.
- Apply the mixed DAB substrate reagent to the tissue section after it has been incubated with the appropriate horseradish peroxidase (HRP) conjugated reagents.
- Approximately 100-200 μl (2-4drops) of the mixed DAB substrate reagent should be sufficient to cover the entire tissue section.
- Incubate the section with the mixed DAB reagent for 2-10 minutes or until the desired color intensity is obtained.
- Wash the slides in dH2O
Warning: DAB is a possible carcinogen. Please take necessary precautions.
Note: This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.