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
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
|
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