User:Floriane Briere/Notebook/CHEM-496/2012/03/20

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Objective

Today's objective is to perform the fluorescence and UV spectroscopy on our samples. We are going to perform our analysis on 6 different samples:

  1. 70 ratio with dye (prepared last 2 weeks)
  2. 166 ratio with dye (prepared last 2 weeks)
  3. BSA/HCl with dye (prepared last 2 weeks)

We are going to use the protocol we established on the 2/22 to 'activate' the dye; so we are going to add Tris buffer until every solution turns blue.

Protocol

  • Preparation of the BSA/HCl control.
  1. Add 1mL of HCl (2.84mM) + 1mL BSA(17.7µM) + 8mL water
  2. Put into the oven for 2 hours
  • Preparation of samples with dye
  1. After dialysis, we obtained:
    1. The 70 ratio with dye was a clear solution with some purple aggregats/beads
    2. The 166 ratio with dye was a clear solution with some purple aggregats/fibers
    3. The BSA/HCl with dye was a pink solution with some pink beads
  2. Add Tris buffer until the solution turns blue
  3. Vortex the samples: the 70 ratio and BSA/HCl solutions dissolved whereas 166 ratio didn't
  4. Centrifugation (5 minutes at 13200rpm) of the 166 ratio with dye solution

Results

  • UV spectroscopy results.

Image:20mar - UV curve.jpg

Image:20mar - UV curve zoom.jpg

  • Fluorescence spectroscopy results.

Image:20mar - fluorescence curve.jpg

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