- Taken from Mary Mendoza.
- Using solutions made last week, a run of 40 μM adenosine with 50 μM varying inhibitors were conducted using the UV 2500 Shimadzu spectrophotometer.
- Three runs of each inhibitor were taken; the average of the runs were taken. The standard deviation were calculated and added as error bars.
- Produce histogram
- The reagents were added in the following sequence:
- 1.78 mL of phosphate buffer
- 600 μL of 200 μM adenosine (final concentration 50 μM)
- This was allowed to mix through venting and placed on the chamber for absorbance reading.
- 300 μL of phosphate buffer (or inhibitor dissolved in phosphate buffer)
- 300 μL of dimethyl sulfoxide (DMSO) (or inhibitor dissolved in DMSO)
- The cuvette contents were mixed by venting and placed on the chamber for absorbance reading.
- 20 μL of adenosine deaminase (ADA) was added to catalyze the reaction
- The kinetics of the reaction was taken using the UVProbe software.
- Cuvette Mixture with Inhibitor
- Cuvette Mixture no Inhibitor
- Solution Preparation/Calculations: Catherine Koenigsknecht
- Cuvette preparation: Mary Mendoza
- Data Evaluation: Dhea Patel
- Overall Lab Assistance: Mike Nagle