User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/11/13: Difference between revisions
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* We can hypothesized that before running the UV-Vis scans for the kinetic assay of ADA with adenosine, adenosine will produce a more pronounced absorption and signal. Therefore, adenosine should be monitored at 235. | * We can hypothesized that before running the UV-Vis scans for the kinetic assay of ADA with adenosine, adenosine will produce a more pronounced absorption and signal. Therefore, adenosine should be monitored at 235. | ||
* Adenosine is the substrate of ADA. As time passes, ADA catalyzes the formation of inosine from adenosine. Thus, the concentration of adenosine should decrease | * Adenosine is the substrate of ADA. As time passes, ADA catalyzes the formation of inosine from adenosine. Thus, the concentration of adenosine should decrease over time. When concentration of adenosine decreases, the absorbance monitored at 235 should also decrease over time. | ||
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ADA Kinetic Assay Preparations
V1 = 0.08934 mL = 89.3 μL in 5 mL of buffer
UV-visible scans of Reagents
M1V1 = M2V2 (3 mM)(10 μL) = M2 (1000 μL) M2 = .0307 mM
3 μL of ADA × 65 μM = M2 (1000 μL) M2 = .195 μM of ADA
M2 = .0975 μM = 97.5 nM
Second dilution M = 48.8 nM Final Concentration = 24.38 nM Beer's Law
[math]\displaystyle{ \frac{\epsilon = A}{bc} }[/math]
For adenosine
For inosine
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