How to Prepare Glucose Oxidase

Glucose oxidase is a commonly used enzyme in analytical applications in which the amount of glucose needs to be determined. Its popularity as a reagent stems from its low cost as compared to other reagents, its stability both dry and in solution, and its low hazard level. Glucose oxidase typically comes in an assay kit that will have most of the reagents you need to prepare the solution with enough materials to perform hundreds of assays. The following steps explain how to prepare the glucose oxidase; your kit will provide specific instructions on how to do the entire assay.

Things You'll Need

• Beakers
• Potassium phosphate (KH2PO4)
• Distilled water
• Stir plate
• pH meter
• 1M Potassium hydroxide solution
• Glucose oxidase
• Test tubes (optional)

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Instructions

• 1

  Prepare a 0.1M potassium phosphate buffer solution by adding 13.6 mg Potassium Phosphate (KH2PO4) into 800 ml of distilled water. Use 1M Potassium Hydroxide to adjust the pH to 7.0 then add enough distilled water to bring the volume to 1 L.

• 2

  Add 10mg of glucose oxidase per 1ml of distilled water in a separate beaker. Mix with gentle shaking and swirling to avoid foaming.

• 3

  Add the glucose oxidase to the 0.1M potassium phosphate buffer in separate beakers or test tubes in the various concentrations needed for your assay. You will need to calculate the concentrations you need based on the detection levels you're trying to analyze and the number of different concentrations you want for your linear regression calculation.