We are running the same procedure as yesterday only with an inhibitor, EHNA.
- 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
- Add 1μL of EHNA to the cuvette and let sit for approximately 1 minute
- 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
- 5mg EHNA in 1mL DMSO ---> 15.9mM EHNA
- (1.9μL)(15.9mM EHNA)=(10mL)(C2). C2 ---> 3μM EHNA
The reaction samples will contain roughly 1nM EHNA
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