User:Jamie Nunziata/Notebook/Protease Research/2015/10/06

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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. 21.2µL of 47.266µM a-chymotrypsin (in phosphate buffer) and 978.8µL of phosphate buffer was added to each sample tube and a blank Eppindorf tube (no fibers), making a final solution of 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


Fluorescence for each sample was measured between the emission spectrum from 400 to 600nm, and the data is recorded below

Data

JMN 10 6 2015 BlankSample Fluorescence.jpg

JMN 10 6 2015 SampleSlope Fluorescence.jpg