User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/08/29

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Contents

Objective

  • to make stock solutions of HAuCl4 and BSA.
  • to make 14 differently concentrated HAuCl4 and BSA mixtures.

Description

  1. Calculations were made to determine how much HAuCl4 was required to make 25mL of 10mM stock solution.
  2. Calculations were made to determine how much BSA was required to make 10mL of 15μM stock solution.
  3. The appropriate amounts of HAuCl4 and BSA were massed separately and added to separate test tubes.
  4. The appropriate amount of H2O were added to each test tube. (25mL to the HAuCl4 test tube and 10mL to the BSA test tube.)
  5. The test tubes were covered with parafilm.
  6. The test tube containing HAuCl4 was also completely covered in aluminum foil.
  7. Then, using the m1v1=m2v2 equation, the amount of each stock solution required to make 4mL mixtures with the Au3+/BSA concentration ratio of 60μM, 80μM, 100μM, 120μM, 128μM, 130μM, 132μM, 133μM, 134μM, 136μM, 138μM, 140μM, 160μM, and 170μM.
  8. After the amounts of each stock solution was determined, the appropriate amounts of HAuCl4, BSA, and H2O were added to each test tube.
  9. The test tubes were capped, covered in aluminum foil, labeled, and placed in the oven at 85°C at 4 hours.

Data and Calculations

  • HAuCl4 Stock
  FW HAuCl4: 393.83 g/mol
  Theoretical Concentration of Stock: 10mM
  Volume of Stock: 25mL
  Theoretical Mass:
        25mL × 10-3 L/mL × 10mM × 10-3 mol/mmol × 393.83 g/mol = 0.0985 g HAuCl4
  Measured Mass: 0.0971 g HAuCl4
  Calculated Concentration: 9862 μM
        0.0971g × (1/393.83 g/mol) × (1/25E-3 L) × 106 μM/mol = 9862 μM 
  • BSA Stock
FW BSA: 66430 g/mol
  Theoretical Concentration of Stock: 15μM
  Volume of Stock: 10mL
  Theoretical Mass:
        10mL × 10-3 L/mL × 15μM × 10-6 mol/mmol × 66430 g/mol = 0.00996 g BSA
  Measured Mass: 0.0105 g BSA
  Calculated Concentration: 15.806 μM
        0.0105g × (1/66430 g/mol) × (1/10E-3 L) × 106 μM/mol = 15.806 μM 
  • Volumes of HAuCl4, BSA, H2 required for various 4mL HAuCl4/BSA mixtures
uM Ratio [Au/BSA] ' Volume [uL] ' '
BSAHAuCl4BSA [μL]HAuCl4H2O [mL]
160253.07147.933.5990
180253.07197.243.5497
1100253.07246.553.5004
1120253.07295.863.4511
1128253.07315.583.4313
1130253.07320.523.4264
1132253.07325.453.4215
1133253.07327.913.4190
1134253.07330.383.4166
1136253.07335.313.4116
1138253.07340.243.4067
1140253.07345.173.4018
1160253.07394.483.3525
1170253.07419.143.3278
4341.75Total uL

Notes

  • The stocks were made with the incorrect concentrations, so the reaction did not occur properly.
  • The stocks were made without the parafilm protection on the spatula.

Image of the Solutions Produced

Image:All.jpg

  • All the solutions from August 29, ranging from 60μM to 170μM Au/BSA concentration. The solution was made without the parafilmed spatula. THe solution spent 4 hours in a 85°C oven.
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