User:Melissa Novy/Notebook/CHEM-571/2012/11/14

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==Entry title==
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==Objectives==
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* Insert content here...
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* Make Au-HRP solutions at mole ratios of Au:HRP from 60 to 150.
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** Analyze the concentration of HRP stock solution with UV-vis.
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** The molar absorption coefficient of HRP at 403 nm is 1.02×10<sup>5</sup> M<sup>-1</sup> cm<sup>-1</sup>.
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* Dialyze fractions of ADA made on x, x, and x.
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==Au-HRP Solutions==
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* Ten solutions at Au-HRP mole ratios of 60, 70, 80, 90, 100, 110, 120, 130, 140, and 150 were made.
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* 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.
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* Note that 130, 140, 150 Au-HRP solutions were recalculated to account for a new stock solution at different concentration of HRP.
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** The concentration of the new HRP stock solution was calculated from its absorbance at 403 nm and its molar absorptivity coefficient.
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==ADA Activity Assay with Adenosine==
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* Frac 1,3 of ADA, made on x, was used.  Its concentration was 10.35 μM, as determined by Bradford assay.
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* It was diluted to 1 ng/μL, then to 0.1 ng/μL using phosphate buffer, prepared on x.
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  '''Initial concentration of ADA: 10.35 μM in 1 mL of solution'''
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  '''(10.35×10<sup>-6</sup>)(0.001 mL) = 1.035×10<sup>-8</sup> mol ADA'''
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  '''(1.035×10<sup>-8</sup> mol)(42000 g/mol) = 0.0004347 g ADA/mL'''
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  '''(0.0004347 g ADA/mL)(1 mL/1000 μL)(10<sup>9</sup> ng/1 g) = 434.7 ng/μL'''
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  '''(434.7 ng/μL)(V<sub>1</sub>) = (1 ng/μL)(1000 μL)'''
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  '''V<sub>1</sub> = 2.3 μL'''
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  '''(1 ng/μL)(V<sub>1</sub>) = (0.1 ng/μL)(1000 μL)'''
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  '''V<sub>1</sub> = 100 μL'''

Revision as of 16:32, 14 November 2012

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Objectives

  • Make Au-HRP solutions at mole ratios of Au:HRP from 60 to 150.
    • Analyze the concentration of HRP stock solution with UV-vis.
    • The molar absorption coefficient of HRP at 403 nm is 1.02×105 M-1 cm-1.
  • Dialyze fractions of ADA made on x, x, and x.

Au-HRP Solutions

  • Ten solutions at Au-HRP 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 H2O added to each solution.
  • Note that 130, 140, 150 Au-HRP 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.

ADA Activity Assay with Adenosine

  • Frac 1,3 of ADA, made on x, was used. Its concentration was 10.35 μM, as determined by Bradford assay.
  • It was diluted to 1 ng/μL, then to 0.1 ng/μL using phosphate buffer, prepared on x.
  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)(109 ng/1 g) = 434.7 ng/μL
  (434.7 ng/μL)(V1) = (1 ng/μL)(1000 μL)
  V1 = 2.3 μL
  (1 ng/μL)(V1) = (0.1 ng/μL)(1000 μL)
  V1 = 100 μL



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