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

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

  • OceanOptics Resultes

Note

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



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