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

Aspirin concentration for ADA Kinetic Assay

• The assay for the concentrations below will be conducted next laboratory period.

ADA Kinetic Assay for obtaining the zero point

• The procedure was taken from 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.

Data