User:Jamie Nunziata/Notebook/Protease Research/2015/11/17

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Objective

To use Ocean Optics to measure absorbance of the degradation of AuNP fibers with 1nM alpha-chymotrypsin over 24 hours.

Protocol

6 fiber samples were spun down at 300rpm for 10 minutes, supernatant was removed, and was carefully brought into solution by 1mL of Tris buffer keeping the fibers as intact as possible. A 1:200 dilution was performed with our 35.156µM a-chymotrypsin stock. 1mL of fiber samples in Tris and 1982.9µL of Tris buffer was added to the cuvette. After the first measurement was collected, 17.1µL of the diluted a-chymotrypsin solution was quickly added to the cuvette between measurements. This final concentration of a-chymotrypsin was 1nM.

Absorbance was collected for 24 hours at various parameters: every 30 seconds for about 1 hour, every 2.5 minutes for about another hour, and every 20 minutes for the remainder of the readings.


Data




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