Most proteins are enzymes, which have an essential role in biological catalysis by increasing the rate of a reaction. The experiment conducted included an enzyme assay with the enzyme catalase and the substrate hydrogen peroxide. To complete the assay, the catalase enzyme was added to the hydrogen peroxide buffered solution. Every thirty seconds, portions from the tube was removed and placed into the labeled tubes. Based on the degree in color of each sample, a different absorption value would be obtained and used in correlation with a working curve of Absorption versus Time (in minutes) to better interpret the catalysis of enzymes in biological catalysis. Introduction
In this experiment, the reaction occurs as the catalase enzyme increases the rate of the decomposition reaction of the substrate to …show more content…
To start, eight empty test tubes were labeled with the corresponding reaction times (in minutes): 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 and additional 3 test tubes labeled “B” (for Blank), “Con” (for controlled), and “Rxn” (for reaction).
2. Then 3 mL of the assay solution was pipetted, using 5 mL pipet, into each test tubes, except for the test tubes labeled “Con” and “Rxn”. The 5 mL pipet was rinsed and kept for later use.
3. Using a 1000 µL, 0.3 mL of the buffer solution was transferred to the test tube labeled “B”, and the test tube was swirled to thoroughly mix the contents.
4. The Enzyme Reaction cocktail was dispensed in 1.8 mL aliquots to each of the “Con” and “Rxn” test tubes using the 5 mL pipet.
5. Afterwards, the “Con” test tube had 0.3 mL of dilute phosphate buffer added to it and the pipet was set aside for later use.
6. 0.3 mL of the contents in the “Con” tube was then added to the tube labeled 0 (assay tube). The “Con” tube was set aside for the rest of the experiment.
7. The test tube labeled “Rxn” had an addition of 0.3 mL of the diluted catalase enzyme placed with a fresh pipet. The tube was thoroughly mixed and the timer was prepared for later
In this experiment, the reaction occurs as the catalase enzyme increases the rate of the decomposition reaction of the substrate to …show more content…
To start, eight empty test tubes were labeled with the corresponding reaction times (in minutes): 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 and additional 3 test tubes labeled “B” (for Blank), “Con” (for controlled), and “Rxn” (for reaction).
2. Then 3 mL of the assay solution was pipetted, using 5 mL pipet, into each test tubes, except for the test tubes labeled “Con” and “Rxn”. The 5 mL pipet was rinsed and kept for later use.
3. Using a 1000 µL, 0.3 mL of the buffer solution was transferred to the test tube labeled “B”, and the test tube was swirled to thoroughly mix the contents.
4. The Enzyme Reaction cocktail was dispensed in 1.8 mL aliquots to each of the “Con” and “Rxn” test tubes using the 5 mL pipet.
5. Afterwards, the “Con” test tube had 0.3 mL of dilute phosphate buffer added to it and the pipet was set aside for later use.
6. 0.3 mL of the contents in the “Con” tube was then added to the tube labeled 0 (assay tube). The “Con” tube was set aside for the rest of the experiment.
7. The test tube labeled “Rxn” had an addition of 0.3 mL of the diluted catalase enzyme placed with a fresh pipet. The tube was thoroughly mixed and the timer was prepared for later