User:Jamie Nunziata/Notebook/Protease Research/2015/11/18

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Objective

Today we used Bradford Assay to redo our run of AuNP degradation from 1nM a-chymotrypsin in hopes of getting better results.


Protocol

6 samples of AuNP fibers were spun down at 300rpm for 10 minutes and the supernatant was removed.These fibers will be used for our 120min, 90min, 60min, 45 min, 30min, and 15min samples. 13.47µL of our 74.22µM stock solution of a-chymotrypsin and 993.17µL of Tris buffer was added to each sample tube and a blank Eppindorf tube (no fibers), making the final solution 1nM a-chymotrypsin.


To each cuvette, the following was added:

  • 1650µL of Tris buffer
  • 750µL of incubated 1µM sample (or blank)
  • 600µL of diluted Bradford Assay in Tris buffer (1:3)

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

JMN 11 18 2015 BlankSamples585 Bradford.jpg



JMN 11 18 2015 CorrectedAborbanceSamples Bradford.jpg



JMN 11 18 2015 Samples585 Bradford.jpg