Objective
- 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
Protocol
- 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 no Inhibitor
- Cuvette Mixture with Inhibitor
- 50uM [2,3-bis(actyloxy)benzoic aicd] [PH003446]
- 50uM [Luteolin] [AMB18511398]
- 50uM [Morin] [AMB18511704]
- 50uM [Robinetin] [AMB18511408]
- 50uM [5-Hydroxy-2-methylbenzoic acid] [Aldrich 696366-1G]
Data
Notes
- Solution Preparation/Calculations: Catherine Koenigsknecht
- Cuvette preparation: Mary Mendoza
- Data Evaluation: Dhea Patel
- Overall Lab Assistance: Mike Nagle
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