User:Dhea Patel/Notebook/CHEM 572: ADA&Inhibitor Kinetics/2013/04/10: Difference between revisions

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==Objective==
==Objective==
* Taken from [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/04/10|Mary Mendoza]].
* Taken from [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/04/10|Mary Mendoza]].
* Using solutions made last week, a run of 40 μM adenosine with 25 μM varying inhibitors were conducted using the UV 2500 Shimadzu spectrophotometer.
* 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.
* 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
* Produce histogram

Revision as of 21:08, 18 April 2013

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Objective

  • 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

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.

Description

  • Cuvette Mixture with Inhibitor

  • Cuvette Mixture no Inhibitor

  • Inhibitors Ran today:
  1. Quercetin
  2. Myricetin

Data

Notes

  • Solution Preparation/Calculations: Catherine Koenigsknecht
  • Cuvette preparation: Mary Mendoza
  • Data Evaluation: Dhea Patel
  • Overall Lab Assistance: Mike Nagle


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