User:Dhea Patel/Notebook/CHEM 572: ADA&Inhibitor Kinetics/2013/02/20

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Aspirin concentration for ADA Kinetic Assay

ADA Kinetic Assay for obtaining the zero point

The procedure is taken from User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/20 Mary Mendoza.
The objective of this laboratory period is to conduct adenosine deaminase (ADA) kinetic assay runs for the new calculated concentrations.
UV 2550 Shimadzu spectrophotometer was baseline with 0.05 M sodium phosphate buffer.
The assays were prepared according to the data below.

After running the first trial, it was observed that the absorbance for 12.34 μM adenosine of trial 2 was superimposed over the 10.52 μM adenosine of trial 1.

It was suggested by Dr. Hartings to use Beer's Law for accurate measurement of the concentration of adenosine in solution.
An article was provided by Dr. Hartings with the molar extinction coefficient, 1.53 x 10^-4 of adenosine at 260 nm.
Thus, it was decided to run a full spectrum of adenosine at each given concentration specified above for accurate measurement of the concentration of adenosine.