User:Beatriz Gimenez De C./Notebook/572/2015/02/03

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Task

  • Data collection of reaction started o Wednesday 28th determine color change
  • Set new protease reaction and monitor periodically

Observations of samples set on the 28th

  • Color change was only present for 57.8 μM pepsin concentration. It took 2 days to be apparent.

Preparation of protease reactions

  • Pepsin
    • Prepare 115.6 μM of pepsin (34600 g/mol) stock solution with 16 mg in 4 ml of H2O
      • 1- 2.5 ml of stock solution + 2.5 mL of buffer ( for a 57.8 μM Pepsin Concentration)
      • 2- 0.5 mL of stock solution + 4.5 buffer (for a 11.56 μM Pepsin Concentration)
      • 3- 0.05 mL of stock solution + 4.95 buffer (for a 1.156 μM Pepsin Concentration)
      • 4- 0.005 mL of stock solution + 5 mL buffer (for a 0.1156 μM Pepsin Concentration)
  • Trypsin
    • Prepare 115.6 μM of trypsin stock solution with 10.77 mg in 4 ml H2O
      • 1- 2.5 ml of stock solution + 2.5 mL of buffer ( for a 57.8 μM Trypsin Concentration)
      • 2- 0.5 mL of stock solution + 4.5 buffer (for a 11.56 μM Trypsin Concentration)
      • 3- 0.05 mL of stock solution + 4.95 buffer (for a 1.156 μM Trypsin Concentration)
      • 4- 0.005 mL of stock solution + 5 mL buffer (for a 0.1156 μM Trypsin Concentration)
  • α-Chymotrypsin
    • Prepare 115.6 μM of α-Chymotrypsin stock solution with 11.56 mg in 4 ml H2O
      • 1- 2.5 ml of stock solution + 2.5 mL of buffer ( for a 57.8 μM α-Chymotrypsin Concentration)
      • 2- 0.5 mL of stock solution + 4.5 buffer (for a 11.56 μM α-Chymotrypsin Concentration)
      • 3- 0.05 mL of stock solution + 4.95 buffer (for a 1.156 μM α-Chymotrypsin Concentration)
      • 4- 0.005 mL of stock solution + 5 mL buffer (for a 0.1156 μM α-Chymotrypsin Concentration)
  • Proteinase K
    • Prepare 115.6 μM of Proteinase K stock solution with 13.37 mg in 4 ml H2O
      • 1- 2.5 ml of stock solution + 2.5 mL of buffer ( for a 57.8 μM Proteinase K Concentration)
      • 2- 0.5 mL of stock solution + 4.5 buffer (for a 11.56 μM Proteinase K Concentration)
      • 3- 0.05 mL of stock solution + 4.95 buffer (for a 1.156 μM Proteinase K Concentration)
      • 4- 0.005 mL of stock solution + 5 mL buffer (for a 0.1156 μM Proteinase K Concentration)

Observations of reactions set today

  • A instantaneous color change was seen with proteinase K at 57.8 μM, at11.56 μM and at 1.156 μM concentrations.


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