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Objective
- Analyze Colloid Dialysis began yesterday
- Perform kinetics experiment described below
- Verify concentrations of CaCl2 solutions used via ISE
Description
- Enzyme Kinetics:
- Use kinetics mode on UV-VIS
- Prepare 10mL 400 unit/mL of lysozyme
- use cold buffer
- you may want to use your 0.6 or 0.12 g/L solution & dilute)
- follow procedure of protocol here.
- Repeat protocol using 4 solutions between 200 & 400 units/mL
- You only need 1 mL of each of these
- Prepare just before addition to keep solution cold
- Set up new dialysis (if time)
Data
Actual Concentrations of Stock CaCl2 Solutions
Ca2+ Stock Solution Concentrations
| Believed CaCl2 Concentration (mM)
|
mV Measurement (mV)
|
Actual CaCl2 Concentration (mM)
|
| 45 |
84.2 |
37.8
|
| 35 |
83.1 |
34.24
|
| 25 |
79.7 |
25.16
|
| 15 |
74.9 |
16.29
|
| 5 |
63.3 |
5.7
|
Ca2+ ISE for Colloid dialzyed against CaCl2
| Original CaCl2 Concentration (mM)
|
mV Measurement (mV)
|
Final CaCl2 Concentration (mM)
|
| 37.8 |
77.8 |
21.19 (bad data point)
|
| 34.24 |
74.8 |
16.14
|
| 25.16 |
70.6 |
11.04
|
| 16.29 |
65.5 |
6.95
|
| 5.7 |
52.6 |
2.16
|
Notes
The colloids crashed out in the 45, 35, 25, and 15 mM CaCl2 dialysis wells from yesterday.
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