User:Jamie Nunziata/Notebook/Protease Research/2015/11/03
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The objective of today's lab was to use a Fluorescence Assay to determine the rate a 10nM solution of a-chymotrypsin degrades AuNP fiber samples.
7 samples of AuNP fibers were spun down at 300rpm for 10 minutes. These fibers will be used for our 24 hour, 120min, 90min, 60min, and 30min samples. A 1:20 dilution of our stock protease was made in phosphate buffer. We pipetted 20µL of our 38.672µM stock solution of a-chymotrypsin into 350µL phosphate buffer. 5.17µL of diluted a-chymotrypsin (in phosphate buffer) and 994.8µL of phosphate buffer was added to each sample tube and a blank Eppindorf tube (no fibers), making a final solution of 10nM a-chymotrypsin.
Fluorescence for each sample was measured between the emission spectrum from 400 to 600nm, and the data is recorded below