User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2014/02/25

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Objective

  • Spin down samples of 30:1 lysozyme-AuNP with 1M salts added and run UV-Vis.
  • Heat samples 30:1 lysozyme-AuNP with 1M salts at 37°C overnight.
  • Image results of 30:1, 55:1, and 110:1 lysozyme-AuNPs synthesized with salt added before heating using UV-Vis.
  • Run atomic absorption of 30:1, 55:1, and 110:1 lysozyme-AuNPs synthesized with salts added before heating.

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.


Atomic Absorption Samples

Solutions with the following Au:lysozyme ratio were run on the AA:

  • 30:1 MgCl2, CaCl2, NaCl, KCl
  • 55:1 MgCl2, CaCl2, NaCl, KCl
  • 110:1 MgCl2, CaCl2, NaCl, KCl


Images

Atomic Absorance of Lysozyme with KCl, NaCl, CaCl2, MgCl2

Figure 1. Gold:protein Ratio to Concentration of Lysozyme-AuNP Baked with Salts

Lysozyme Gold-protein to Concentration with Salts, JAC.png

UV-Vis

UV-Vis For Solutions that Contained Salt Before Lysozyme AuNP Synthesis

Screen shot 2014-02-25 at 11.48.04 PM.png Screen shot 2014-02-25 at 11.47.52 PM.png Screen shot 2014-02-25 at 11.47.42 PM.png Screen shot 2014-02-25 at 11.47.31 PM.png

UV-Vis For Select Variables at More Refined Concentrations

  • Please Note that the Control Wasn't Diluted With Variable Like the Other Samples.

Screen shot 2014-02-25 at 11.47.14 PM.png Screen shot 2014-02-25 at 11.46.46 PM.png Screen shot 2014-02-25 at 11.46.17 PM.png Screen shot 2014-02-25 at 11.45.14 PM.png Screen shot 2014-02-25 at 11.45.00 PM.png Screen shot 2014-02-25 at 11.44.28 PM.png Screen shot 2014-02-25 at 11.44.01 PM.png

  • Based off of all UV-Vis data: The ABS for the controls were corrected for the dilution factor present and the percent AuNP's in solution was calculated.

Screen Shot 2014-03-26 at 2.49.43 PM.png

Notes

Notes.2.25.jac.png

  • These results detail the samples from last Thursday with the addition of the variables to 1 mL aliquots of 30:1 ratio with salts added after.


More.2.25.notes.png

  • These results detail the reaction of the different ratios synthesized with salts added before heating.