User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/11/14

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Revision as of 15:09, 9 December 2012

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During the previous lab entry, the molar absorptivities of adenosine and inosine were calculated. Adenosine was found to have a higher molar absorptivity at 235.
It was concluded from the previous period that the absorption of adenosine should decrease over time. A kinetic assay scan of the reaction containing 2.7 mL of 0.05 M sodium phosphate buffer, 300 μL of 3.07 mM adenosine, and 15 μL of 42 μM ADA was taken.
The program was set to Kinetics with the wavelength at 235 for a collection span of 600s.

In the scan, it can be observed that the absorption of adenosine is not decreasing over time. Instead, the absorption is increasing. Also, the signal for the absorption of adenosine is over 1.
It was decided that the next measurements, the concentration of adenosine should be decreased by decreasing the volume in th cuvette. This is to ensure that the absorbance is below 1.

Volume of ADA (μL)	Concentration of ADA	Volume of phosphate buffer (mL)	Concentration of phosphate buffer (M)	Volume of adenosine (μL)	Concentration of adenosine (mM)
15		2.95	0.051	50	0.051
50		2.95	0.051	50	0.051

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 ==ADA Kinetic Assay Runs==
 * During the previous lab entry, the molar absorptivities of adenosine and inosine were calculated. Adenosine was found to have a higher molar absorptivity at 235.
-* It was concluded from the previous period that the absorption of adenosine should decrease over time. A kinetic assay scan of the reaction containing 2.7 mL of sodium phosphate buffer, 300 μL of adenosine, and 15 μL of ADA was taken.
+* It was concluded from the previous period that the absorption of adenosine should decrease over time. A kinetic assay scan of the reaction containing 2.7 mL of 0.05 M sodium phosphate buffer, 300 μL of 3.07 mM adenosine, and 15 μL of 42 μM ADA was taken.
 * The program was set to Kinetics with the wavelength at 235 for a collection span of 600s.