Difference between revisions of "Peroxidase UV Vis assay"
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| − | + | The focus of our research is the activity of sea urchin ovoperoxidase. We are starting with Horse Radish Peroxidase (HRP) to develop a standard protocol for measuring the activity of a peroxidase. | |
==Standard Mixing Procedure== | ==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 mixture of Peroxidase UV-Vis assays requires the use of a Guaiacol stock solution and a hydrogen peroxide stock solution. | ||
Revision as of 01:46, 12 November 2018
The focus of our research is the activity of sea urchin ovoperoxidase. We are starting with Horse Radish Peroxidase (HRP) to develop a standard protocol for measuring the activity of a peroxidase.
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.