User:Roberta Diaz Jimenez/Notebook/CHEM 471 Experimental Lab/2015/09/29

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Perform a Bradford Analysis to analyze the degradation of protease trypsin.


In order to do the Bradford Assay of Protease Degradation, we followed Dr. Hartings' protocol, and did the following specific steps:

  • Protease Sample Prep.
  1. Used eppendorf tube no. 1 that weighed 1.0269g, and contained 0.00108g.
  2. Added 1mL of phosphate/CaCl2 buffer.
  3. Final concentration: 46.35µM
  • Sample Prep.
  1. Used 7 eppendorf tubes, each containing gold fibers.
  2. To each tube add:
    1. 0.9784mL of buffer
    2. 0.02157mL of trypsin (add this at the time of putting the tubes in the hot water bath).
  • Blank Prep.
  1. In on eppendorf tube add:
    1. 0.9784mL of buffer
    2. 0.02157mL of trypsin
  • Bradford Dilution
  1. Dilute 1mL of Bradford in 3mL of buffer such that the resulting solution is 1:4.


   V1 = [(1µM)(1mL)]/46.35µM = 0.02157mL, amount of trypsin solution needed
   Volume of buffer: 1mL - 0.02157mL = 0.9784mL


Graph 1. BradfordGraph1Sept29.png Graph 2. BradfordGraph2Sept29.png