User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2015/03/18

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10 nM Chymotrypsin kinetics

Another in situ kinetics run. 104.1 µL of the dilute chymotrypsin stock was diluted with 2.896 mL of buffer for a final concentration of 10 nM when added to the fibers. The fibers and OceanOptics were prepared with the same protocol as previous kinetics runs.

Results

10nM Chymotrypsin AbsvsTime Mar18 Chart.png

There was no visible colorimetric change in the solution. Fibers were still present at the end of the allotted reaction time.

Protease Bradford - Control

250 μL samples of protease solutions in Tris/CaCl2 buffer at the experimental concentrations (1 nM, 10 nM, 100 nM, and 1 μM) were combined with 200 mL of 1:4 (Bradford:Buffer) Bradford reagent and 550 mL of additional Tris/CaCl2 buffer in order to determine the contribution of the protease protein to the Bradford absorbances observed in the AuNP fiber samples.

Results

Control Proteinase K Bradford Absorbance.png

Control Trypsin Bradford Absorbance.png

Control Chymotrypsin Bradford Absorbance.png

Control Thermolysin Bradford Absorbance.png