User:Jamie Nunziata/Notebook/Protease Research/2015/11/11: 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 1nM a-chymotrypsin protease | ||
==Procedure== | |||
7 samples of AuNP fibers were spun down at 300rpm for 10 minutes.These fibers will be used for our 23 hours, 120min, 90min, 60min, and 30min samples. A 1:400 diluion of our stock protease was made in Tris buffer. We pipetted 2.5µL of our 58.60µM stock solution of a-chymotrypsin into 997.5µL of Tris buffer. 6.83µL of the diluted a-chymotrypsin (in Tris) 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 [[User:Nicole_Bonan/Notebook/Chem_571_Lab_Notebook/2015/11/04|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_11_11_2015_CorrectedAborbanceSamples_Bradford.jpg]] | |||
[[Image:JMN_11_11_2015_BlankSample600nm_Bradford.jpg]] | |||
[[Image:JMN_11_11_2015_Sample600nm_Bradford.jpg]] | |||
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Revision as of 20:58, 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 1nM a-chymotrypsin protease Procedure7 samples of AuNP fibers were spun down at 300rpm for 10 minutes.These fibers will be used for our 23 hours, 120min, 90min, 60min, and 30min samples. A 1:400 diluion of our stock protease was made in Tris buffer. We pipetted 2.5µL of our 58.60µM stock solution of a-chymotrypsin into 997.5µL of Tris buffer. 6.83µL of the diluted a-chymotrypsin (in Tris) 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.
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 11 11 2015 CorrectedAborbanceSamples Bradford.jpg
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