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

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Objective

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.

Description

  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)

Results

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

5oct-fluoIconc=f(WL).png

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