User:Jamie Nunziata/Notebook/Protease Research/2015/11/10
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The objective of today's lab was to use a Fluorescence Assay to determine the rate a 1nM solution of a-chymotrypsin degrades AuNP fiber samples.
6 samples of AuNP fibers were spun down at 300rpm for 10 minutes. These fibers will be used for our 48 hours, 24 hours, 120min, 90min, 60min, and 30min samples. A 1:200 dilution of our stock protease was made in phosphate buffer. We pipetted 5µL of our 51.172µM stock solution of a-chymotrypsin into 995µL phosphate buffer. 3.91µL of diluted a-chymotrypsin (in phosphate buffer) and 996.1µL of phosphate buffer was added to each sample tube and a blank Eppindorf tube (no fibers), making a final solution of 1nM a-chymotrypsin.
Fluorescence for each sample was measured between the emission spectrum from 400 to 600nm, and the data is recorded below