User:Khyra A. Neal/Notebook/Chem 571/2014/10/28

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

October 28, 2014

Tasklist

1. Dialysis Extraction of 0.6 g/L Lysozyme vs KI prepared on October 22, 2014

  • Extract sample solutions from dialysis chamber and store in 30 mL extraction vials


2. Bradford Analysis of Extracted Lysozyme and KI solutions

  • 200 μL Bradford Reagent (diluted 1:3)
  • 50 μL Sample Solution
  • 750 μL 50 mM Tris/ 50 mM NaCl Buffer
  • Run UV-Vis between 400 nm and 800 nm

Bradford Lys vs KI.jpg


3. UV-Vis of Extracted Lysozyme Solutions vs KI and Fluorescence

  • Dilute Lysozyme samples 1:100
  • Add 100 μL of Sample to small volume cuvette
  • Run UV-Vis between 200 nm- 800 nm
  • Clean cuvette in between samples with 1 N HCl, 1% SDS solution, and Methanol
    • There was no need to run fluorescence because KI is interfering with the ability of Bradford to bind to the lysozyme protein.


4. I- ISE

KI concentration mV measurement (mV) (21 Oct.) mV measurement (mV) (28 Oct.)
2mM -198.3 -192.8
5 mM -217.3 -206.8
10 mM -237.0 -224.2
25 mM -247.4 -253.0
50mM -278.0 -276.2


5. Dialysis Preparation

  • Prepare dialysis chamber for 0.6 g/L Lysozyme vs CaCl2
  • Add 1 mL of 0.6 g/L Lysozyme solution to each well on one side of the chamber
  • Add 1 mL of CaCl2 to the opposite side of the chamber
    • 5 μM CaCl2
    • 50 μM CaCl2
    • 500 μM CaCl2
    • 5 mM CaCl2,
    • 50 mM CaCl2
  • Sit on low speed shaker overnight and prepare for analysis