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
- Mix 100μL of 2.9mM HAuCl4 and 100μL 17.7μM BSA with 800μL distilled water in a microcentrifuge tube.
- Place the tube on a heat block at 80°C for three hours.
- Add 500μL 50mM Tris, pH 7.5.
- Remove from heat after about 5 min. Observe and take a spectra.
Studying the Fibers and Solutions from Yesterday
- Centrifuge the microcentrifuge tube with purple protein fibers from yesterday (ratio=163.842) for 5 min at 13200rpm.
- Take a spectrum of the solution (without disturbing the pellet).
- 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.
A spectrum was taken of this solution, which showed a peak at 534nm:
Studying the Fibers and Solutions from Yesterday
Spectra were taken of different solutions from yesterday:
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:
Tube Number |
Ratio [Au]/[BSA] |
Wavelength Peak
|
1
|
163.842 |
544nm
|
2
|
131.073 |
533nm
|
5
|
65.5367 |
535nm
|
6
|
32.7684 |
N/A
|
|