User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2012/02/21

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Objective

To take UV-Vis and Fluorescence of the dialysis samples and to make additional BSA control.

Description

  1. A BSA control solution was prepared by mixing 8 mL of water, 1 mL of 2.84 mM HCl, and 1 mL of 17.7 μM BSA.
  2. A control solution of dye in water was prepared by dissolving 1 mg of dye in 80 uL of water and then adding 5 mL water.
  3. The reaction mixtures were transferred from dialysis tubing to plastic containers.
  4. 2 1 mL samples of both the 70 (Gold solution) and 166 (Purple Fibers) molar ratio reactions were placed in a microcentrifuge for 5 minutes at 13200 rpm.
  5. UV-Vis spectra were taken between 200 and 800 nm of 6 samples (BSA control with dye, 70 molar ratio with dye, 166 molar ratio with dye, 70 ratio without dye, and 166 ratio without dye)
  6. Fluorescence spectra were taken of the BSA control with dye at excitations of 525, 550, 575, and 600 nm. It was determined that 600 nm was the best wavelength for excitation and therefore was used in the following trials: BSA control with dye, 70 molar ratio with dye, and 166 molar ratio with dye.


Data

UV-Vis Results

Image:21feb - UV curve.jpg

Image:21feb - UV curve zoom.jpg

Fluorescence Results

Image:21fev - Fluorescence curve.jpg

Notes

  • After dialysis, the BSA/HCl control was a pink solution, the 70 molar ratio was had a pink solution with small purple fibers, and the 166 molar ratio had a pink solution with a large purple aggregate.
  • A peak was expected to be seen in the mid 600s in the Fluorescence spectra, because the dye emits at 672 nm. This peak was not observed, likely because the pH of the solutions was under 8.00.


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