User:Helen L. Slucher/Notebook/CHEM 571/2013/10/08

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Objective

Observe and measure ADA turnover kinetics in the absence of an inhibitor.

Description

  1. Make a 40μM solution of adenosine in buffer (50mM phosphate buffer, pH 7.4)
  2. Go to Dr. Hartings lab for enzyme kinetics measurements.
    1. Add 3mL of adenosine solution to the cuvette
    2. Start your kinetics measurement
      1. 1ms integration (on front panel)
      2. 10 scan average (on front panel)
      3. Set "Save the first available scan every" to 15 seconds (after clicking File>Save)
      4. Set "Stop after this amount of time" to 10 minutes (after clicking File>Save)
      5. Set "File Type" to Tab Delimited
      6. Give the files a directory and a name
      7. Click accept
      8. Just before 1 minute add 30ul of 0.01u/mL ADA

Data

  • Screen shot 2013-10-29 at 4.15.29 PM.png

Notes

  • Stock ADA = 3140 uM
  1. 0.0042 g adenosine
  2. 5 mL buffer
  • 40 uM ADA
  1. 0.13 mL stock
  2. 9.87 mL buffer