User:Beatriz Gimenez De C./Notebook/572/2015/03/18

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Tasks

  • Measure the mean residue liquid in the fiber sample
    • 115 μL, 175 μL and 340 μL for a mean of 210 μL
  • Measure the absorbance of the different protease concentrations used
    • 250 μL samples of protease solutions in Tris/CaCl2 buffer at 1 nM, 10 nM, 100 nM, and 1 μM
    • Add 200 mL of 1:4 (Bradford:Buffer) Bradford + 550 mL of additional Tris/CaCl2 b
  • 10 nM Chymotrypsin kinetics
    • 104.1 µL of the dilute chymotrypsin stock was diluted + 2.896 mL of buffer --> 10 nM
    • OceanOptics (used same protocol as previous kinetics runs)

Results

  • Protease Bradford results

Control Proteinase K Bradford Absorbance.png

Control Trypsin Bradford Absorbance.png

Control Chymotrypsin Bradford Absorbance.png

Control Thermolysin Bradford Absorbance.png

  • AuNP Protease Corrected Bradford Spectra
    • Proteinase K

Proteinase K Bradford 1uM Corrected.png

Proteinase K Bradford 100 nM Corrected.png

Proteinase K Bradford 10nM Corrected.png

Proteinase K Bradford 1nM Corrected.png

    • Trypsin

Trypsin Bradford 1uM Corrected.png

Trypsin Bradford 100 nM Corrected.png

Trypsin Bradford 10nM Corrected.png

Trypsin Bradford 1nM Corrected.png

    • Chymotrypsin

Chymotrypsin Bradford 1uM Corrected.png

Chymotrypsin Bradford 100nM Corrected.png

Chymotrypsin Bradford 10nM Corrected.png

Chymotrypsin Bradford 1nM Corrected.png

    • Thermolysin

Thermolysin Bradford 1uM Corrected.png

Thermolysin Bradford 100nM Corrected.png

Thermolysin Bradford 10nM Corrected.png

Thermolysin Bradford 1nM Corrected.png

  • OceanOptics Resultes

10nM Chymotrypsin AbsvsTime Mar18 Chart.png

Note

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