User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/18: Difference between revisions
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==Procedure== | ==Procedure== | ||
* For details on making stock solutions, see [[User:Melissa Novy/Notebook/CHEM-571/2012/09/18|Melissa's Notebook]] for the final concentration of stock solution for horseradish peroxidase, sodium phosphate buffer, 4-Aminoantipyrine, and 4-Iodophenol. | * For details on making stock solutions, see [[User:Melissa Novy/Notebook/CHEM-571/2012/09/18|Melissa's Notebook]] for the final concentration of stock solution and final concentration reached for horseradish peroxidase, sodium phosphate buffer, 4-Aminoantipyrine, and 4-Iodophenol. | ||
* Samples mixed with altering concentration of H2O2 was made and analyzed under UV-vis spectrometer. The concentration of AAP(4-aminoantipyrine) was set at 2.5mM, 4-Iodophenol was set at 18mM, and horseradish peroxidase at 2.3uM. The concentration of H2O2 was changed from 1.7mM to 106.25uM. | * Samples mixed with altering concentration of H2O2 was made and analyzed under UV-vis spectrometer. The concentration of AAP(4-aminoantipyrine) was set at 2.5mM, 4-Iodophenol was set at 18mM, and horseradish peroxidase at 2.3uM. The concentration of H2O2 was changed from 1.7mM to 106.25uM. | ||
* In the next set of samples, concentration of H2O2 is held constant. The concentration of AAP was altered from 1.25mM to 156.25uM. The concentration of other solutions were kept the same. | * In the next set of samples, concentration of H2O2 is held constant. The concentration of AAP was altered from 1.25mM to 156.25uM. The concentration of other solutions were kept the same. | ||
* For results obtained from UV-vis spectrometer, see [[User:Melissa Novy/Notebook/CHEM-571/2012/09/18|Melissa's Notebook]] for graph of Absorbance versus Time of enzymatic activity of HRP in Fixed [AAP] and varied [H2O2], and in Fixed [H2O2] and varied [AAP] | * For results obtained from UV-vis spectrometer, see [[User:Melissa Novy/Notebook/CHEM-571/2012/09/18|Melissa's Notebook]] for graph of Absorbance versus Time of enzymatic activity of HRP in Fixed [AAP] and varied [H2O2], and in Fixed [H2O2] and varied [AAP]. In those samples, the initial and final concentration reached were depicted in the table below: | ||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Stock Solution''' | |||
| align="center" style="background:#f0f0f0;"|'''Initial Concentration, M<sub>1</sub> [M]''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume Added V<sub>1</sub> [mL]''' | |||
| align="center" style="background:#f0f0f0;"|'''Final Concentration, M<sub>2</sub> [M]''' | |||
|- | |||
| H<sub>2</sub>O<sub>2</sub>||0.000034||0.75||0.000017 | |||
|- | |||
| AAP||0.0025||0.7||0.001166667 | |||
|- | |||
| Phenol||0.018||0.01||0.00012 | |||
|- | |||
|} | |||
==Notes== | ==Notes== | ||
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* It was discovered that when concentration of H2O2 is held constant, the enzymatic activity increases as the concentration of AAP increases. The results can be shown in [[User:Melissa Novy/Notebook/CHEM-571/2012/09/18|Melissa's Notebook]] in graph Absorbance over time of Enzymatic Activity of HRP in Fixed [H2O2] and Varied [AAP] by comparing the slopes of each sample run. | * It was discovered that when concentration of H2O2 is held constant, the enzymatic activity increases as the concentration of AAP increases. The results can be shown in [[User:Melissa Novy/Notebook/CHEM-571/2012/09/18|Melissa's Notebook]] in graph Absorbance over time of Enzymatic Activity of HRP in Fixed [H2O2] and Varied [AAP] by comparing the slopes of each sample run. | ||
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Revision as of 20:19, 26 October 2012
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