October 28, 2014
Primary Experiment #1 - KI Analysis
Because the titrations done last week demonstrated inconsistent results, the KI concentrations of each of the ten dialysis samples were re-analyzed using the self-made Iodide Ion Selective Electrode. The results are as follows:
Sample
|
Potential (mV)
|
ln[KI]
|
[KI] (mM)
|
Sample
|
Potential (mV)
|
ln[KI]
|
[KI] (mM)
|
L+50 mM KI |
-253.7 |
3.014259251 |
20.37399334 |
50 mM KI |
N/A |
N/A |
N/A |
|
L+25 mM KI |
-237.7 |
2.324990307 |
10.22658096 |
25 mM KI |
-240.1 |
2.428380649 |
11.34050295
|
L+10 mM KI |
-215.8 |
1.38155344 |
3.981081197 |
10 mM KI |
-210.8 |
1.166156895 |
3.209633945
|
L+5 mM KI |
-199.9 |
0.696592427 |
2.006902376 |
5 mM KI |
-196.2 |
0.537198983 |
1.711207024
|
L+2 mM KI |
-177.4 |
-0.272692026 |
0.761327221 |
2 mM KI |
-173.7 |
-0.432085469 |
0.649153892
|
NOTE: No more 50 mM KI sample remained to test with I- ISE due to an error that occurred during the titration.
Primary Experiment #3 - Work-up
0.6 g/L Lysozyme vs CaCl2 using MWCO 3500
Well 1
|
Well 2
|
Well 3
|
Well 4
|
Well 5
|
0.6 g/L lysozyme
|
0.6 g/L lysozyme
|
0.6 g/L lysozyme
|
0.6 g/L lysozyme
|
0.6 g/L lysozome
|
50 mM CaCl2
|
5 mM CaCl2
|
0.5 mM CaCl2
|
50 μM CaCl2
|
5 μM CaCl2
|
Necessary Analyses:
Bradford
- 5 lysozyme samples, 0.6 g/L lysozyme reference, and Bradford blank
- 50 μL sample, 200 μL 1:4 Bradford, 750 μL 50 mM Tris/50 mM NaCl
Fluorescence
- 5 lysozyme samples, 0.6 g/L lysozyme reference
- 10 μL of sample in 990 μL of DI water
UV-Vis
- 5 lysozyme samples, 0.6 g/L lysozyme reference
- 10 μL of sample in 990 μL of DI water
NOTE: The 5 µM sample was re-run because the wavelength range was not correct the first time the sample(s) was/were run. A lysozyme stock reference sample and Bradford/Tris/NaCl blank were re-run for comparison as well.
CaCl2 ISE
Concentration
|
Lysozyme Solution Potential (mV)
|
Soak Solution Potential (mV)
|
50 mM |
88.6 |
87.8
|
5 mM |
61.3 |
63.5
|
0.5 mM |
30.0 |
39.0
|
50 µM |
10.5 |
16.3
|
5 µM |
8.9 |
26.0
|
Primary Experiment #2 - Set-up Take 2
30:1 colloid solution vs CaCl2 using MWCO 3500
NOTE: Because the results of the Bradford analysis of this dialysis (take 1) were ridiculous, for lack of a better word, this dialysis was re-run and re-analyzed.
Well 1
|
Well 2
|
Well 3
|
Well 4
|
Well 5
|
30:1 colloid
|
30:1 colloid
|
30:1 colloid
|
30:1 colloid
|
30:1 colloid
|
50 mM CaCl2
|
5 mM CaCl2
|
0.5 mM CaCl2
|
50 μM CaCl2
|
5 μM CaCl2
|
|