User:Jamie Nunziata/Notebook/Protease Research/2015/10/13

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Objective

The objective of today's lab was to use a Fluorescence Assay to determine the rate a 1µM solution of a-chymotrypsin degrades AuNP fiber samples.

Procedure

7 samples of AuNP fibers were spun down at 300rpm for 10 minutes.These fibers will be used for our 120min, 90min, 60min, 45min, 30min, 15min, and 10min samples. 21.0µL of 47.656µM a-chymotrypsin (in Tris) and 979.0µL of Tris buffer was added to each sample tube and a blank Eppindorf tube (no fibers), making the final solution 1µM a-chymotrypsin.


To each cuvette, the following was added:

  • 20uL of the blank or sample
  • 140uL of Assay Buffer
  • 40uL of Assay Reagent

A full lab procedure can be found in Nicole Bonan's notebook entry for today


Absorbance for each cuvette was measure via UV-Vis between the emission sprectrum of 400-800nm, and the data is recorded below


Data



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