User:Carly M. Montanero/Notebook/CHEM-571/2014/03/05

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Biomaterials Design Lab: Spring 2014 Main project page
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Objective

  • Calculate equilibrium constants for heated 30:1 lysozyme:AuNP (with variables added after baking) and room temperature 30:1 lysozyme:AuNP (with variables added after baking).
  • Make standard with amount of gold per 5 mL tube to run AA and UV-Vis.
  • Centrifuge samples from yesterday (30:1 lysozyme-AuNP with variables added after heating at room temperature).
  • Run AA and UV-Vis on the centrifuged samples.

Procedure

Atomic Absorption

Creating the Gold Stock Solutions

  1. Add 50 μL of HAuCl4·3H2O and 4950 μL of distilled water to a Falcon tube, for a final concentration of 10 μg/mL Au.
  2. Add 100 μL of HAuCl4·3H2O and 4900 μL of distilled water to a Falcon tube, for a final concentration of 20 μg/mL Au.
  3. Add 150 μL of HAuCl4·3H2O and 4850 μL of distilled water to a Falcon tube, for a final concentration of 30 μg/mL Au.
  4. Add 200 μL of HAuCl4·3H2O and 4800 μL of distilled water to a Falcon tube, for a final concentration of 40 μg/mL Au.
  5. Add 250 μL of HAuCl4·3H2O and 4750 μL of distilled water to a Falcon tube, for a final concentration of 50 μg/mL Au.
  6. A sixth stock solution of gold was created with 448 μL of HAuCl4·3H2O and 4552 μL of distilled water to a Falcon tube. This resulting concentration represents the amount of gold in a 5 mL sample of lysozyme-AuNP.


Atomic Absorption Samples

The following samples were run on the AA:

  • 10 μg/mL Au, 20 μg/mL Au, 30 μg/mL Au, 40 μg/mL Au, 50 μg/mL Au
  • 30:1 lysozyme-AuNP with NaCl, KCl, MgCl2, CaCl2, 2,2 bipyridine, MES, citric acid
  • 60:1, 70:1, 80:1, 90:1, 100:1 BSA-AuNP with MgCl2
  • Gold solution in 5 mL sample of lysozyme-AuNP
  • Control of 30:1 lysozyme-AuNP without any added variable

Figures

Figure 1. Atomic absorbance of Lysozyme with different concentrations of variables


Figure 2. Atomic Absorbance of BSA with MgCl2 added before being heated

Image:BSA with MgCL2 Javier Vinals Camallonga.png


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