|
Objective
- Analyze dialysis solutions from yesterday
- UV-Vis
- Fluoresence
- Bradford
- Calcium ion ISE
- Measure dialysis concentrations from last week
- Prepare a new dialysis using colloid solution and CaCl2 or HCl
- For those running CaCl2, use 3500 MWCO & test concentrations between 10 and 50 mM
- For those running HCl, use 25000 MWCO & test concentrations between 100 μM and 10 mM
- Prepare solutions for enzyme kinetics
- Prepare 25 mL 66 mM Potassium phosphate, pH 6.24 (beaker prep is fine)
- Place half in a Falcon tube, label & refrigerate overnight
- Prepare 10 mL 0.01 wt% Micrococcus lysodeikticus Cell Suspension (in freezer) using phosphate buffer
- Adjust the cell suspension concentration until the Absorbance at λ = 450 nm is between 0.6 & 0.7
Data
Ca2+ ISE for Lysozyme dialzyed against CaCl2
| Original CaCl2 Concentration (mM)
|
mV Measurement (mV)
|
Final CaCl2 Concentration (mM)
|
| 6.83 |
55.0 |
2.68
|
| 1.87 |
39.9 |
0.684
|
| 0.68 |
2.9 |
0.024
|
| 0.020 |
-5.7 |
0.011
|
| 0.00058 |
-0.3 |
0.0179 (This is a bad data point)
|
New Dialysis of Colloid
Colloid was dialyzed against CaCl2 of the following concentrations
- 45 mM
- 35 mM
- 25 mM
- 15 mM
- 5 mM
|
Biomaterials Design Lab
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