Objectives
 Make AuADA solutions at mole ratios of Au:ADA from 60 to 150.
 Make AuHRP solutions at mole ratios of Au:HRP from 60 to 150.
 Analyze the concentration of HRP stock solution with UVvis.
 The molar absorptivity coefficient of HRP at 403 nm is 1.02×10^{5} M^{1} cm^{1}.
 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 Dhea Patel's entry for theoretical and actual calculations of stock solutions, and volumes of stock solutions and deionized H_{2}O added to each solution.
 Note that ADA made on 2012/10/03 was used to make the solutions. The solution of HAuCl_{4} was made on User:Melissa_Novy/Notebook/CHEM571/2012/09/05.
AuHRP Solutions
 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_{2}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.
 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^{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
 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)(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
References
R&D Systems ADA Activity Assay
AU Biomaterials Design Lab ADA Activity Assay
