Objective
- Run all tests on the Lysozyme-CaCl2 dialysis
- Make more lysozyme colloid solution
- Set up a dialysis of Lysozyme-CaCl2 with new concentrations
Description
- Colloid Synthesis
- make 0.04621g HAuCl4 / 50mL H2O
- make 0.0448g Lysozyme / 25mL H2O
- make a 30:1 HAuCl4 Lysozyme solution
- 0.5mL HAuCl4, 4.17mL Lysozyme, and 0.33mL 50mL H2O
- Heat in an oven over 4 hours at 80°C
- Its go time - 0.6g Lysozyme against 50mM, 5mM, 500μM, 50μM, 5μM CaCl2
- Bradford Analysis
- 50μL sample
- 200μL Bradford reagent
- 4:1 DI Water to Bradford Assay
- 750μL Tris-NaCl Buffer
- UV Protein Analysis
- Fluorescence Analysis
- ISC of CaCl2
- The probe was allowed to sit in solution before taking measurements, this seemed to yield better results. These measurements may need to be taken again to double check
Data
- Bradford Analysis
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'
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y=0.0206x+0.0969
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Abs. at 590nM |
Concentration of Lysozyme (ug/mL)
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Bradford Blank |
0.204 |
5.199029126
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5uM CaCl2 |
0.46 |
17.62621359
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50uM CaCl2 |
0.467 |
17.96601942
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500uM CaCl2 |
0.431 |
16.2184466
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5mM CaCl2 |
0.466 |
17.91747573
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50mM CaCl2 |
0.43 |
16.16990291
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- Table Displaying Calculated Concentrations based off of a Calibration curve of absorbance against a known concentration of Lysozyme.
- UV Protein Analysis
- Fluorescence Analysis
- ISC of CaCl2
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ISC - CaCl2
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Potential=26.344(Concentration)+40.946
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Theoretical Concentration (mM) |
Potential (mV) |
|
log (concentration) |
Actual Concentration (mM)
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Samples |
50.000 |
89.7 |
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1.850668084 |
70.90356695
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5.000 |
65.6 |
|
0.93584877 |
8.62678094
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0.500 |
46 |
|
0.191846341 |
1.555415207
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|
0.050 |
32.1 |
|
-0.335788035 |
0.461542783
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|
0.005 |
26.1 |
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-0.563543881 |
0.273184541
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Stocks |
50.000 |
93.8 |
|
2.006301245 |
101.4614922
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5.000 |
65.9 |
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0.947236562 |
8.855978681
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0.500 |
43.8 |
|
0.108335864 |
1.283322663
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0.050 |
28.5 |
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-0.472441543 |
0.336944566
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|
0.005 |
15 |
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-0.984892196 |
0.103539915
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Notes
Always discuss methods between groups. It can only help you perfect your procedures.
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