Objective
- Prepare reagents for horseradish peroxidase (HRP) assay
- Complete HRP assay with and without gold nanoparticles to see if they interact with HRP's active site
Procedure
- HRP procedure File:3760-Protocol HRP AbsorbanceAssay.pdf was followed. Time was plotted against absorbance on the UV/Vis spectroscoper.
- 1mL horseradish peroxidase was dissolved in 1mL distilled water.
- A .2M buffer of sodium phosphate was made at a pH of 7.
- .0017M hydrogen peroxide solution was made by adding 1mL 30% hydrogen peroxide to 100mL distilled water. 1mL of hydrogen peroxide solution was diluted to 50mL with 49mL of sodium phosphate buffer for 34μM solution.
- 698μM iodophenol solution was made in 1mL DMSO, since it's not soluble in water. 2.5mM 4-aminoantipyrine solution was prepared with the sodium phosphate buffer.
Discussion
- The HRP assays were not completed today, but will be 10/02, with the reagents prepared today
|