User:Michael S. Bible/Notebook/571/2014/11/11

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Objective

  • Analyze Dialysis Started on 11/5
    • UV-Vis of Protein solutions
    • KI ISE of Ligand solutions
    • Fluorescence Analysis of Protein Solutions
    • Bradford Analysis of Protein Solutions

Bradford Analysis

Bradford was performed using:

  • 1:4 Bio-Rad Solution
  • 50 μL of protein solutions
  • 200 μL of Bradford Reagent
  • 750 μL of 50 mM NaCl/Tris Buffer

Bradford Analysis of KI Dialyzed Lysozyme.png

UV Analysis of KI Dialyzed Lysozyme

UV Analysis of KI Dialyzed Lysozyme.png

KI ISE Data

Original KI Concentration (mM) mV Measurement (mV) Final KI Concentration (mM)
66.65 -253.4 16.79
39.41 -243.8 11.21
19.69 -239.3 9.28
11.45 -220.6 4.23
3.23 -193.5 1.35

Fluorescence

Fluorescence was run from λ = 300 - 550 nm and at an excitation wavelength of λ=280 nm.

Fluorescence of KI Dialyzed Lysozyme.png

New Dialysis Experiment

A new dialysis was set up using:

  • 0.6 g/L
  • 50 mM, 5 mM, 500 μM, 50 μM, 5 μM CaCl2