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

Biomaterials Design Lab: Spring 2014

Main project page Previous entry      Next entry

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 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. 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. 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. 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. 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. 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