User:Floriane Briere/Notebook/CHEM-496/2012/03/21

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Objective

According to yesterday's results, we haven't been able to reproduce the fluorescence results we obtained on the 2/22 whereas the UV results are consistent. In fact, on the 2/22 and on the 3/20 we obtained for the UV spectroscopy a curve where the BSA/HCl (with dye) is the highest, then it's the 70 ratio (with dye) and the lowest is the 166 ratio (with dye). However, for the fluorescence spectroscopy we obtained different results. On the 2/22, the highest were the 70 ratio (with dye) and the BSA/HCl (with dye); these two results were very closed to each other; the lowest was the 166 ratio. On the 3/20, the highest was the 166 ratio (with dye); the lowest were the 70 ratio (with dye) and BSA/HCl (with dye); these two results were very closed to each other. So, results are only partially reproducible. Today we are going to redo the measurements on yesterday's solutions to check our results then, we are going to figure out what could have happened and see if we can reproduce the 2/22 results.

Protocol

  • Spectroscopy measurements with same solutions as yesterday.
  1. Centrifugation (5 minutes at 13200rpm) for:
    1. 166 ratio with dye solution
    2. 70 ratio with dye solution
    3. BSA/HCl with dye solution
  2. Take fluorescence spectrum
  • Spectroscopy measurements after having changed the pH of solutions.
  1. Measure the pH:
    1. 166 ratio solution with dye: pH = 8.13
    2. 70 ratio solution with dye: initial pH = 7.62
    3. BSA/HCl solution with dye: initial pH = 7.75
  2. Add Tris buffer:
    1. 70 ratio solution with dye: added 3 drops; final pH = 8.07
    2. BSA/HCl solution with dye: added 3 drops; final pH = 8.09
  3. Centrifugation (3 solutions): 5 minutes at 13200rpm
  4. Take fluorescence spectrum

Results

  • Fluorescence spectroscopy results (same solutions as yesterday).

  • Fluorescence spectroscopy results (after having changed the pH of the solutions).

  • UV spectroscopy results (after having changed the pH of the solutions).

Notes

This area is for any observations or conclusions that you would like to note.


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