User:Megan L. Channell/Notebook/Horseradish/2013/10/08

Biomaterials Design Lab

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Objective Measure and observe ADA kinetics without inhibitor. This work will be the basis of our comparison to ADA-AuNPtunrover studies. From Dr. Hartings. Procedure Make a 40μM solution of adenosine in buffer (50mM phosphate buffer, pH 7.4) Go to Dr. Hartings lab for enzyme kinetics measurements. Add 3mL of adenosine solution to the cuvette Start your kinetics measurement 1ms integration (on front panel) 10 scan average (on front panel) Set "Save the first available scan every" to 15 seconds (after clicking File>Save) Set "Stop after this amount of time" to 10 minutes (after clicking File>Save) Set "File Type" to Tab Delimited Give the files a directory and a name Click accept Just before 1 minute add 30ul of 0.01u/mL ADA taken from Dr. Hartings How we made the 40μM solution of adenosine in buffer .1063 grams of adenosine were dissolved in a 10mL volumetric flask. Molarity: ~40mM the solution was mixed to ensure homogenity 9.98 μL of 40mM solution were diluted up to 10mL. Molarity: 40μM Data Notes Adenosine deaminase (ADA) stock 1.1mg (24.0 units in 1 mg) in 25mL of buffer --> 1.1 units/mL ADA ADA for experiments 100uL of stock ADA and 900uL buffer --> 0.11 units/mL ADA