User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/12: Difference between revisions

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==ADA Kinetic Assay==
*The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05|02/05/2013]], calculated values.
*The objective of performing the assay is to obtain  the rate of the enzymatic reaction of adenosine into inosine by ADA.
*Baseline 200-400nm using pH 7.4, 0.05M phosphate buffer
*Temperature control: 25°C
*Phosphate buffer in reference cell
*Ran kinetics on 250uM solution (20uL ADA, 600uL 1250uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
**absorbance exceeded 4.00, so the assay was stopped and a new solution was prepared.
*Ran kinetics on 2.56uM solution (20uL ADA, 600uL 12.8uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
**dAbs was obtained and plotted vs. time in excel
*Ran kinetics on 6.4uM solution (20uL ADA, 600uL 32uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
**dAbs was obtained and plotted vs. time in excel
*Ran kinetics on 16uM solution (20uL ADA, 600uL 80uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
**dAbs was obtained and plotted vs. time in excel
*The following table describes the volumes and concentrations of the components added to the cuvette.




[[Image:Data_Table.JPG|center]]




Revision as of 15:15, 8 May 2013

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