User:Khyra A. Neal/Notebook/Chem 571/2014/10/14: Difference between revisions
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** Run UV-Vis of undialyzed 0.6 g/L Lysozyme solution with Bradford reagent | ** Run UV-Vis of undialyzed 0.6 g/L Lysozyme solution with Bradford reagent | ||
[[Image: Dialyzed_Lysozyme_in_KI.jpg]] | |||
3. Calibration Curve for undialyzed Lysozyme by Bradford Analysis | 3. Calibration Curve for undialyzed Lysozyme by Bradford Analysis | ||
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* Measure UV-Vis from 400 nm - 800 nm using polystyrene cuvettes. | * Measure UV-Vis from 400 nm - 800 nm using polystyrene cuvettes. | ||
[[Image: Undialyzed_Lysozyme_Calibration.jpg]] | |||
4. Prepare 50 mL 66 mM Potassium Phosphate Buffer | 4. Prepare 50 mL 66 mM Potassium Phosphate Buffer | ||
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* Transfer 100 μL to a small volume fluorescence cuvette | * Transfer 100 μL to a small volume fluorescence cuvette | ||
* Run fluorescence from 300 nm -550 nm and excitation at 280 nm | * Run fluorescence from 300 nm -550 nm and excitation at 280 nm | ||
6. UV-Vis | |||
* Run Uv-Vis on protein containing solutions that were dialyzed on [http://openwetware.org/wiki/User:Khyra_A._Neal/Notebook/Chem_571/2014/10/08 October 8, 2014] | |||
* Run from 200 nm -400 nm | |||
7. Prepare New Dialysis for 30:1 Colloid vs CaCl<sub>2</sub> | |||
* Use 3500 MWCO tubing | |||
* Add 1 mL 30:1 colloid solution to 5 cells on one side of chamber | |||
* Add 1 mL 5 μM, 50 μM, 500 μM, 5 mM, and 50 mM CaCl<sub>2</sub> to the opposite side of the chamber (one concentration per cell) | |||
* Secure wells by screwing them to prevent any evaporation | |||
* Place on low speed shaker overnight | |||
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Latest revision as of 00:26, 27 September 2017
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October 14, 2014Tasklist1. SDS Page #2
3. Calibration Curve for undialyzed Lysozyme by Bradford Analysis
4. Prepare 50 mL 66 mM Potassium Phosphate Buffer
5. Fluorescence on protein containing solutions that were dialyzed on October 8, 2014
6. UV-Vis
7. Prepare New Dialysis for 30:1 Colloid vs CaCl2
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