User:Jamie Nunziata/Notebook/Protease Research/2015/09/30

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Objective

The purpose of this lab is to use Bradford Assay to measure AuNP fiber degradation from a solution of 1µM a-chymotrypsin protease

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. 24.5µL of 40.625µM a-chymotrypsin (in Tris) and 975.4µL 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:

  • 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

Image:JMN_9_30_2015_CorrectedAborbanceSample_Bradford.jpg

Image:JMN_9_30_2015_BlankSample600nm_Bradford.jpg

Image:JMN_9_30_2015_CorrectedSample600nm_Bradford.jpg



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