User:Floriane Briere/Notebook/CHEM-496/2011/10/05

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Today's experiment consists on taking fluorescence spectrum of 4 different MPB solutions. This is going to provide us information about the MPB fluorescing property.


  1. Group 1 prepared 4 different MPB solutions:
    1. Solution A: 1397µL of MPB + 1603µL of Tris buffer
    2. Solution B: 300µL of solution A + 2700µL of Tris buffer
    3. Solution C: 300µL of solution B + 2700µL of Tris buffer
    4. Solution D: 300µL of solution C + 2700µL of Tris buffer
  2. Performed a fluorescence spectrum for each solution (excitation at 450nm; emission range from 500 to 700nm)


This experiment was performed to determine the fluorescing property of MPB; the MPB solution is the same as in the Bradford Assay. The first step of this experiment was to determine the sensitivity of the instrument by using standard solutions of [Ru(bpy)2-(5aminophen)](PF6)2. Then, we measured the fluorescence of five different MPB solutions; each solution is characterized by a specific concentration. The five different emission spectra were performed with an excitation at 450nm and an emission range from 500 to 700nm. To finish, we took a spectrum at 280nm of the five different solutions.

  • Ratio of intensity/concentration as a function of the wavelength


Using the molar absorptivity of MPB which has been determined thanks to the Bradford Assay, we have been able to calculate the concentration of the five MPB solutions: -Solution A : 1.09*10-5 M -Solution B : 1.08*10-6 M -Solution C : 1.67*10-7 M -Solution D : 4.18*10-8 M -Solution E : 2.09*10-8 M