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Objectives

• Filter the cell protein solution made on 2012/10/02 to obtain ADA using fast protein liquid chromatography (FPLC).
• Make 10 mL of Au/BSA solutions at the usual mole ratios for atomic absorption.

Au/BSA Solutions for AA

• Due to a miscalculation, the stock solutions for HAuCl4 and for BSA were combined. The total number of moles of BSA and moles of HAuCl4 in this new solution were calculated and then new volumes of BSA stock solution were determined:
 Stock Solution Concentrations [M] ' [HAuCl4] 0.026 [BSA] 0.000003

• There are 0.00000208 mol of HAuCl4 per 0.00008 L of solution.
• There are 2.4E-10 mol of BSA per 0.00008 L of solution.
• Please refer to the following table for calculations and concentrations:
 Au:BSA Mol BSA Needed Mol BSA Added Volume BSA Added [L] Volume BSA Added [uL] 60 3.46667E-08 3.44267E-08 0.002295111 2295.111111 80 0.000000026 2.576E-08 0.001717333 1717.333333 100 2.08E-08 2.056E-08 0.001370667 1370.666667 120 1.73333E-08 1.70933E-08 0.001139556 1139.555556 128 1.625E-08 1.601E-08 0.001067333 1067.333333 130 0.000000016 1.576E-08 0.001050667 1050.666667 132 1.57576E-08 1.55176E-08 0.001034505 1034.505051 133 1.56391E-08 1.53991E-08 0.001026607 1026.606516 134 1.55224E-08 1.52824E-08 0.001018826 1018.825871 136 1.52941E-08 1.50541E-08 0.001003608 1003.607843 138 1.50725E-08 1.48325E-08 0.000988831 988.8309179 140 1.48571E-08 1.46171E-08 0.000974476 974.4761905 160 0.000000013 1.276E-08 0.000850667 850.6666667 170 1.22353E-08 1.19953E-08 0.000799686 799.6862745

• The protocol for making the AA Au/BSA solutions is as follows:
1. Add 80 μL of Au/BSA stock solution to each test tube.
2. Then add the given volumes of BSA stock solution to each test tube (refer to Volume BSA Added [μL]).
3. Dilute all solutions to a final volume of 10 mL.
4. Place the solutions in an 80°C oven for 4 hours.