Difference between revisions of "User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/04/19"

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==Objective==
 
* To obtain a spectrum of 40 μM adenosine and inosine
 
* 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
 
  
==Procedure for Adenosine and Inosine Spectrum==
 
* To obtain spectrum of 40 μM adenosine, dilute 600 μL of 200 μM adenosine stock solution in 2400 μL 0.05 M phosphate buffer.
 
* Stock solution of 200 μM inosine (MW 268.2 g/mol) was prepared. A spectrum was taken using the same dilution mentioned above.
 
  
==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
 
[[Image:CuvetteMixtureNoInhibitor.png]]
 
  
*Cuvette Mixture with Inhibitor
 
[[Image:CuvetteSolnInhibitorHisto.png]]
 
  
*Inhibitors Ran today:
 
  
 
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Revision as of 14:45, 8 May 2013

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