Medico DAB Substrate (3, 3’-diaminobenzidine) produces a brown reaction product in the presence of peroxidase (HRP) enzyme. Substrate Kit contains all of the reagents necessary toprepare. These reagents are supplied in convenient dropper bottles.

REAGENTS

STORAGE

• Store reagents in original bottles at the required conditions (one year)

• Avoid storing reagents or working solution in strong direct light.

PREPARATION OF SUBSTRATE WORKING SOLUTION

• To 1.0 ml of distilled water

  • Add 50 µl of Reagents C

  • Add 50 µl of Reagents A

  • Add 50 µl of Reagents B

• Mix well before use

INSTRUCTIONS FOR USE

For Tissues or Cells After incubation with a peroxidase (HRP) detection system, rinse well. Incubate with the substrate working solution at room temperature for 2-10 minutes. Optimal development times should be determined by the investigator. Wash for 5 minutes in water.

NOTES

• We recommend using glass-distilled water in the preparation of substrate buffer. Deionized water may con-tain inhibitors of the peroxidase reaction. Solutions containing sodium azide or other inhibitors of peroxidase activity should not be used in diluting the peroxidase substrate.

• DAB are carcinogens. Appropriate care should be exercised when using these reagents including gloves, eye protection, lab coats, and good laboratory procedures.

Content

Size

Storage

Reagents A

DAB   20×concentrated solution

3ml

-20℃

Reagents B

H2O2  20×concentrated solution

3ml

4-8℃

Reagents C

TBS   20× concentrated solution

3ml

4-8℃