User:Tamra L. Fisher/Notebook/Experimental Biological Chem/2011/12/07

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Objective

To see if the formation of purple protein fibers can be prevented when removing from heat if a refolding buffer is added first. Also, to see if the solution with fibers from yesterday also has purple solution.

Description

Studying the Effect of Refolding Buffer

  1. Mix 100μL of 2.9mM HAuCl4 and 100μL 17.7μM BSA with 800μL distilled water in a microcentrifuge tube.
  2. Place the tube on a heat block at 80°C for three hours.
  3. Add 500μL 50mM Tris, pH 7.5.
  4. Remove from heat after about 5 min. Observe and take a spectra.

Studying the Fibers and Solutions from Yesterday

  1. Centrifuge the microcentrifuge tube with purple protein fibers from yesterday (ratio=163.842) for 5 min at 13200rpm.
  2. Take a spectrum of the solution (without disturbing the pellet).
  3. Take spectra of other solutions from yesterday.

Data

Studying the Effect of Refolding Buffer

Picture of the microcentrifuge tube with the Au/BSA solution and refolding buffer. No fibers formed when the tube was removed from heat.

Photo (11).JPG


A spectrum was taken of this solution, which showed a peak at 534nm:

Spectra refolding 120711.png

Studying the Fibers and Solutions from Yesterday

Spectra were taken of different solutions from yesterday:

Specta 120711.png


A zoomed-in spectra shows a shift in peak. The solution that had fibers in it (highest ratio) has a peak around 544nm, and the next highest ratio has a peak shifted to about 533nm. From there the peak barely shifts. The solution with the lowest ratio did not show any purple or pink peaks:

Zoomed spectra 120711.png


Tube Number Ratio [Au]/[BSA] Wavelength Peak
1 163.842 544nm
2 131.073 533nm
5 65.5367 535nm
6 32.7684 N/A