User:Khyra A. Neal/Notebook/Chem 571/2014/10/22

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Ocotober 22, 2014

Tasklist

  • Extract Dialyzed 30:1 Colloid Solutions against CaCl2 that were dialyzed on October 21,2014

Bradford Analysis of Extracted Colloid Solutions

  • 100 μL sample
  • 200 μL Bradford Reagent diluted (1:3)
  • 700 μL 50 mM Tris/ 50 mM NaCl buffer
  • Run UV-Vis between 400 nm and 800 nm

Image:Bradford_of_Colloid_in_Cacl2.jpg

3. UV-Vis and Fluorescence

There are no peaks at 280 nm for the dialyzed colloid solutions because phenylalanine, tryptophan, and tyrosine are reducing agents to gold. There is no free protein remaining in solution after dialysis.

The diagrams below confirm the above statement.

  • Insert Diagram here*


4. Prepare dialysis of 0.6 g/L Lysozyme vs KI

  • Use 3500 MWCO tubing
  • Add 1 mL 0.6 g/L Lysozyme solution to 5 cells on one side of chamber
  • Add 1 mL 2 mM, 5 mM, 10 mM, 25 mM, and 50 mM KI to the opposite side of the chamber (one concentration per cell)
  • Secure wells by screwing them to prevent any evaporation
  • Place on low speed shaker for one week
    • Our lysozyme stock solution precipitated out so we are using 0.6 g/ L Lysozyme that was prepared by Monika today. O


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