User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/20

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ADA Kinetic Assay for obtaining the zero point

  • 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.
Screen Shot 2013-02-19 at 2.46.05 PM.png
  • 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.