User:Melissa Novy/Notebook/CHEM571/2012/11/14
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** The molar absorptivity coefficient of HRP at 403 nm is 1.02×10<sup>5</sup> M<sup>1</sup> cm<sup>1</sup>.  ** The molar absorptivity coefficient of HRP at 403 nm is 1.02×10<sup>5</sup> M<sup>1</sup> cm<sup>1</sup>.  
* Dialyze fractions of ADA made on x, x, and x.  * Dialyze fractions of ADA made on x, x, and x.  
+  
+  ==AuADA Solutions==  
+  * Ten solutions at AuADA mole ratios of 60, 70, 80, 90, 100, 110, 120, 130, 140, and 150 were made.  
+  * Please refer to [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/14Dhea Patel's entry]] for theoretical and actual calculations of stock solutions, and volumes of stock solutions and deionized H<sub>2</sub>O added to each solution.  
+  * Note that ADA made on [[User:Melissa_Novy/Notebook/CHEM571/2012/10/032012/10/03]] was used to make the solutions.  
==AuHRP Solutions==  ==AuHRP Solutions==  
* Ten solutions at AuHRP mole ratios of 60, 70, 80, 90, 100, 110, 120, 130, 140, and 150 were made.  * Ten solutions at AuHRP mole ratios of 60, 70, 80, 90, 100, 110, 120, 130, 140, and 150 were made.  
  * Please refer to Dhea Patel's entry for theoretical and actual calculations of stock solutions, and volumes of stock solutions and deionized H<sub>2</sub>O added to each solution.  +  * Please refer to [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/14Dhea Patel's entry]] for theoretical and actual calculations of stock solutions, and volumes of stock solutions and deionized H<sub>2</sub>O added to each solution. 
* Note that 130, 140, 150 AuHRP solutions were recalculated to account for a new stock solution at different concentration of HRP.  * Note that 130, 140, 150 AuHRP solutions were recalculated to account for a new stock solution at different concentration of HRP.  
** The concentration of the new HRP stock solution was calculated from its absorbance at 403 nm and its molar absorptivity coefficient.  ** The concentration of the new HRP stock solution was calculated from its absorbance at 403 nm and its molar absorptivity coefficient.  
+  ** Its concentration was calculated to be 28.5 μM.  
+  
+  '''Abs.: 2.1'''  
+  '''Path length (b): 1 cm'''  
+  '''Molar absorptivity coefficient (ε) = 1.02×10<sup>5</sup> M<sup>1</sup> cm<sup>1</sup>'''  
+  '''Concentration (c) = 2.1/(1 cm)(1.02×10<sup>5</sup> M<sup>1</sup> cm<sup>1</sup>) = 28.5 μM'''  
==ADA Activity Assay with Adenosine==  ==ADA Activity Assay with Adenosine== 
Revision as of 16:45, 14 November 2012
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Objectives
AuADA Solutions
AuHRP Solutions
Abs.: 2.1 Path length (b): 1 cm Molar absorptivity coefficient (ε) = 1.02×10^{5} M^{1} cm^{1} Concentration (c) = 2.1/(1 cm)(1.02×10^{5} M^{1} cm^{1}) = 28.5 μM ADA Activity Assay with Adenosine
Initial concentration of ADA: 10.35 μM in 1 mL of solution (10.35×10^{6})(0.001 mL) = 1.035×10^{8} mol ADA (1.035×10^{8} mol)(42000 g/mol) = 0.0004347 g ADA/mL (0.0004347 g ADA/mL)(1 mL/1000 μL)(10^{9} ng/1 g) = 434.7 ng/μL (434.7 ng/μL)(V_{1}) = (1 ng/μL)(1000 μL) V_{1} = 2.3 μL (1 ng/μL)(V_{1}) = (0.1 ng/μL)(1000 μL) V_{1} = 100 μL ReferencesR&D Systems ADA Activity Assay
