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* 30:1 lysozyme-gold colloid
* 30:1 lysozyme-gold colloid
** 1 mL 250 mM HAuCl<sub>4</sub> , 2 mL 42 μM (0.6 g/L) Lysozyme, and 7 mL HPLC-grade H<sub>2</sub>O
** Add 0.5 mL 250 mM HAuCl<sub>4</sub> , 1 mL 42 μM (0.6 g/L) Lysozyme, and 3.5 mL HPLC-grade H<sub>2</sub>O to 5 culture tubes
** Place in oven for 4 hours
** Place in oven for 4 hours



Latest revision as of 00:26, 27 September 2017

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October 7, 2014

Tasklist

1. Solution Preparation

  • 42 μM = 0.6 g/L Lysozyme
    • add 0.015 g Lysozyme to 25 mL deionized water
  • 30:1 lysozyme-gold colloid
    • Add 0.5 mL 250 mM HAuCl4 , 1 mL 42 μM (0.6 g/L) Lysozyme, and 3.5 mL HPLC-grade H2O to 5 culture tubes
    • Place in oven for 4 hours
  • 50 mm NaCl 50 mM Tris buffer solution was made by adding:
    • 0.3030g of Tris buffer
    • 0.1461g of NaCl
    • Add to 50 mL distilled water


2. Bradford Analysis

  • Solutions were prepared on October 1, 2014
    • 20 μL sample solution
    • 200 μL Bradford Reagent (diluted 1:3 with Tris buffer)
    • 780 μL 50 mM Tris/ 50 mM NaCl Buffer
      • Solutions were vortexed in a 1.5 mL centrifuge tube and equilibrated for 5 minutes
      • Ran UV-Vis between 400-800 nm using PS cuvettes
      • An Undialyzed Lysozyme Bradford analysis was also ran.
  • Transfer remaining dialyzed solutions to an extraction vial.

3. pH Measurements of H+ containing solutions

pH measurements

Sample pH
0.25 mM HCl (4) 8.062
Lysozyme (4) 8.134

4. Ca2+ ISE of Ca2+ containing solutions

Substance Potential (mV)
Colloid Sample 1 73.5
50 mM CaCl2 (1) 74.9
Colloid Sample 2 51.2
500 μM CaCl2 91.4