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

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

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

JMN 9 30 2015 CorrectedAborbanceSample Bradford.jpg

JMN 9 30 2015 BlankSample600nm Bradford.jpg

JMN 9 30 2015 CorrectedSample600nm Bradford.jpg