User:Nancy T. Miles/Notebook/N.Miles Lab 2013-09-03/2013/10/09

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Objective

We are running the same procedure as yesterday only with an inhibitor, EHNA.

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. Add 1μL of EHNA to the cuvette and let sit for approximately 1 minute
    3. 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

EHNA stock

  1. 5mg EHNA in 1mL DMSO ---> 15.9mM EHNA
  2. (1.9μL)(15.9mM EHNA)=(10mL)(C2). C2 ---> 3μM EHNA

The reaction samples will contain roughly 1nM EHNA


Notes

This area is for any observations or conclusions that you would like to note.


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