Assay Lab Report

ABSTRACT The relationship between the amount of substrate in the assay solution and the rate of the reaction when the enzyme and buffer in the assay are held constant were experimented. We analyzed the change in absorbencies over time for varying substrate concentrations. There were four experimental assays which contained 1% enzyme solution, substrate solution of 0%, 1%, 2%, and 3% concentrations, guaiacol, and pH 7 buffer. At 2% concentration there was a greater enzymatic activity and at 3% concentration enzymatic activity decreased. The results were supported of our hypothesis.
INTRODUCTION
Enzymes are proteins that catalyze chemical reactions and are one of the most important proteins in the human body. Enzymes work by lowering activation energy,
The 0% concentration of Hydrogen Peroxide was used as our control. The 3% concentration of Hydrogen Peroxide was provided so all that was needed to do was prepare the 0%, 1%, and 2% concentrations which was done by using the equation C₁V₁ = C₂V₂. Other materials consisted of pH 7 buffer, guaiacol which was used as our indicator, and 1% turnip peroxidase solution. Eight cuvettes were needed, four of which were blank tubes which means it did not contain guaiacol, the indicator. The other four cuvettes were experimental tubes. The pH 7 buffer, peroxidase, guaiacol, and Hydrogen Peroxide were added into the experimental tubes. The order in which you add these solutions is very important. The Hydrogen Peroxide must be added last and guaiacol should be added one tube at a time. When ready to put into the spectrophotometer the blank tubes should be put in first to blank the machine for a more accurate absorbance reading when the experimental tubes are put in. The absorbances for the experimental tubes were recorded every fifteen seconds for two minutes. This process was done for the remaining cuvettes.