Peroxidase UV Vis assay

introduction...We are interested in measuring the peroxiase activity of a solution. We are going to isolate ovop from sea urchins and we need to know how actie the enzyme is. We are starting with HRP and ...

Standard Mixing Procedure

The mixture of Peroxidase UV-Vis assays requires the use of a Guaiacol stock solution and a hydrogen peroxide stock solution. The stock solutions are prepared as follows:

Guaiacol Stock Solution

49.5 microliters of Guaiacol is added to a 25 mL pH 8.0 buffer solution, which is comprised of 25 mL of DI water and a 8.0 pH buffer tablet that has been mixed thoroughly.

Hydrogen Peroxide Stock Solution

17 microliters of hydrogen peroxide is added to 1 mL of DI water.

Assay

5.0 mL Guaiacol stock solution is added to a vial.

10 microliters of the hydrogen peroxide stock solution is added to the Guaiacol stock solution.

The desired amount of peroxidase is then added to the assay before the UV-Vis is ran.