User:Jamie Nunziata/Notebook/Protease Research/2015/10/14: Difference between revisions
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== | ==Objective== | ||
* | The purpose of this lab is to use Bradford Assay to measure AuNP fiber degradation from a solution of 100nM 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. 2.44µL of 41.016µM a-chymotrypsin (in Tris) and 997.6µL 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 [[User:Nicole_Bonan/Notebook/Chem_571_Lab_Notebook/2015/10/14|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_10_14_2015_CorrectedAborbanceSample_Bradford.jpg]] | |||
[[Image:JMN_10_14_2015_BlankSample600nm_Bradford.jpg]] | |||
[[Image:JMN_10_14_2015_CorrectedSample600nm_Bradford.jpg]] | |||
Revision as of 20:16, 24 November 2015
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ObjectiveThe purpose of this lab is to use Bradford Assay to measure AuNP fiber degradation from a solution of 100nM a-chymotrypsin protease Procedure7 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. 2.44µL of 41.016µM a-chymotrypsin (in Tris) and 997.6µL 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:
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 DataFile:JMN 10 14 2015 CorrectedAborbanceSample Bradford.jpg
File:JMN 10 14 2015 CorrectedSample600nm Bradford.jpg
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