User:Monika Gasiorek/Notebook/CHEM-571 2014F/2014/09/16: Difference between revisions
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==Dialysis== | ==Dialysis== | ||
5 dialysis experiments were set up: | |||
I) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM NaCl stock solution. | |||
II) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM glycine stock solution. | |||
III) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of pure water. | |||
IV) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 25k g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM glycine stock solution. | |||
V) 4.0 mL of the 35.6:1 [Au]:[lys] solution wad dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM glycine stock solution. | |||
NOTE: A sixth dialysis using 15k g/mol MWCO tubing was intended (4.0 mL of lysozyme solution against 50 mM glycine soln), but the specific tubing was not available. | |||
==Calibration Curves: pH and Conductivity== | ==Calibration Curves: pH and Conductivity== |
Revision as of 18:29, 16 September 2014
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September 16, 2014Stock Solution PreparationA 500 mL stock solution of 50 mM NaCl was made.
NOTE: The purpose of this solution was to serve as a dialyzing agent A 1 L stock buffer solution of 50 mM glycine with a pH of approx. 3.5 (pKa in glycine buffer range) was made.
NOTE: The purpose of this solution was also to serve as a dialyzing agent. A buffer solution was necessary in order to change the ionic strength of the solution without altering its pH.
A 25 mL solution of 1 g/L lysozyme was made.
NOTE: The purpose of this solution was to serve as a baseline for the measurement of the equilibria/movement of free protein and/or ions across the dialysis tubing. A 10.0 mL sample of approximately 30:1 (rather 36.5:1) gold:protein colloid solution was obtained from previous experiments.
Dialysis5 dialysis experiments were set up: I) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM NaCl stock solution. II) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM glycine stock solution. III) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of pure water. IV) 4.0 mL of the prepared lysozyme solution was dialyzed in about 3 in of 25k g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM glycine stock solution. V) 4.0 mL of the 35.6:1 [Au]:[lys] solution wad dialyzed in about 3 in of 3500 g/mol MWCO tubing in a 150 mL beaker filled with approx. 120 mL of the 50 mM glycine stock solution. NOTE: A sixth dialysis using 15k g/mol MWCO tubing was intended (4.0 mL of lysozyme solution against 50 mM glycine soln), but the specific tubing was not available. Calibration Curves: pH and Conductivity |