User:Monika Gasiorek/Notebook/CHEM-571 2014F/2015/03/18

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March 18th, 2015

Protease Bradford - Control

250 μL samples of protease solutions in Tris/CaCl2 buffer at the experimental concentrations (1 nM, 10 nM, 100 nM, and 1 μM) were combined with 200 mL of 1:4 (Bradford:Buffer) Bradford reagent and 550 mL of additional Tris/CaCl2 buffer in order to determine the contribution of the protease protein to the Bradford absorbances observed in the AuNP fiber samples.

Results

Image:Control_Proteinase_K_Bradford_Absorbance.png

Image:Control_Trypsin_Bradford_Absorbance.png

Image:Control_Chymotrypsin_Bradford_Absorbance.png

Image:Control_Thermolysin_Bradford_Absorbance.png

AuNP Protease Corrected Bradford Spectra

Proteinase K

Image:Proteinase_K_Bradford_1uM_Corrected.png

Image:Proteinase_K_Bradford_100_nM_Corrected.png

Image:Proteinase_K_Bradford_10nM_Corrected.png

Image:Proteinase_K_Bradford_1nM_Corrected.png

Trypsin

Image:Trypsin_Bradford_1uM_Corrected.png

Image:Trypsin_Bradford_100_nM_Corrected.png

Image:Trypsin_Bradford_10nM_Corrected.png

Image:Trypsin_Bradford_1nM_Corrected.png

Chymotrypsin

Image:Chymotrypsin_Bradford_1uM_Corrected.png

Image:Chymotrypsin_Bradford_100nM_Corrected.png

Image:Chymotrypsin_Bradford_10nM_Corrected.png

Image:Chymotrypsin_Bradford_1nM_Corrected.png

Thermolysin

Image:Thermolysin_Bradford_1uM_Corrected.png

Image:Thermolysin_Bradford_100nM_Corrected.png

Image:Thermolysin_Bradford_10nM_Corrected.png

Image:Thermolysin_Bradford_1nM_Corrected.png

10 nM Chymotrypsin kinetics

Another in situ kinetics run. 104.1 µL of the dilute chymotrypsin stock was diluted with 2.896 mL of buffer for a final concentration of 10 nM when added to the fibers. The fibers and OceanOptics were prepared with the same protocol as previous kinetics runs.

Results

There was no visible colorimetric change in the solution. Fibers were still present at the end of the allotted reaction time.



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