Difference between revisions of "User:Michael F. Nagle/Notebook/Chem 571/2012/11/13"

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==Goals==
 
==Goals==
 
* Analyze kinetics of [[AU_Biomaterials_Design_Lab:Materials/ADA Protein| Adenosine Deaminase ]] with UV/Vis
 
* Analyze kinetics of [[AU_Biomaterials_Design_Lab:Materials/ADA Protein| Adenosine Deaminase ]] with UV/Vis
* Analyze samples of various mole ratios of HRP to [[AU_Biomaterials_Design_Lab:Materials/HAuCl4|HAuCl<sub>4</sub>]] prepared [[http://openwetware.org/wiki/User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/07|11/07 by Dhea Patel]]
+
* Analyze samples of various mole ratios of HRP to [[AU_Biomaterials_Design_Lab:Materials/HAuCl4|HAuCl<sub>4</sub>]] prepared [[http://openwetware.org/wiki/User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/07|11/07 by Dhea Patel]]
  
 
==Procedure==
 
==Procedure==

Revision as of 04:24, 16 November 2012

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Goals

Procedure

  1. Dhea Patel measured the absorbance of HRP/Au samples with UV/Vis
  2. The procedure for an ADA Activity Assay was followed
    1. The buffer used was made with sodium phosphate rather than potassium phosphate.
  1. Puja Moody prepared the buffer at a pH of 7.4. The sodium and potassium phosphate buffers are interchangeable since they share the phosphate ion.
  1. Samples of Inosine, Adenosine and ADA in buffer were analyzed via UV/Vis
    1. Samples of Inosine, Adenosine and ADA in buffer were analyzed via UV/Vis
    2. Absorbance for adenosine maxed out the instrument, so the .1mM solution was further diluted with buffer to .5mM, which had absorbance near 5.
    3. Since this was still close to maxing out the machine, the adenosine was diluted to .05mM.
    4. Absorbance for 65μM ADA maxed out the instrument, so a serial dilution was completed
      1. 500μL ADA solution was transferred to 500μL sodium phosphate buffer, and 500μL of the resulting solution was transferred to 500μL. The transfer was repeated three times, producing solutions at the concentrations .195μM, 97.5μM, 48.75nM, and 24.38nM.

Data

Adacharts.png