User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2015/02/18
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10 nM Protease K Kinetics Run
Determine the kinetics of Protease K on AuNP fibers at 10 nM enzyme concentration
69 μL of the 0.4325 μM stock was diluted with 2.931 mL of Tris/NaCl buffer. This was added onto the fibers in the cuvette. The reaction was run at 37°C for 6 hours, scanning every 5 minutes.
The reaction did have a discernible colorimetric change. There were still fibers present in solution after 6 hours. A longer reaction may see this go to completion.
Determination of Trypsin Cleavage of AuNP Fibers
1. 0.151 mL of stock solution in 4.849 mL of Tris/NaCl buffer
2. 0.015(1) mL of stock solution in 4.985 mL of Tris/NaCl buffer
3. 0.001(5) mL of stock solution in 4.998 mL of Tris/NaCl buffer
4. 10 μL of stock solution in 990 μL of Tris/NaCl buffer --> [trypsin] = 0.3305 μM
Fibers were spun down at 300 RPM for 5 minutes, and liquid extracted. 5 mL of protease solution was added to each sample tube and a 500 μL aliquot from each concentration was obtained every 30 minutes beginning with time point 0. The samples were left in the incubator shaker (37C) for 3.0 hours.