Objective
- Using UV/Vis, find which mole ratios of HAuCl4 to BSA result in the most BSA unfolding and formation of gold nanoparticles.
- Make Tris stock solutions and do serial dilution with HAuCl4 and BSA fibers in Tris buffer
Procedure
- UV/Vis
- 1mL of each solution with a varying mole ratios of HAuCl4 and BSA was inserted into a cuvette, which went into a UV/Vis Spectroscoper
- Spectra were obtained and peaks for gold nanoparticles were identified at the 520nm range.
- Cuvette was cleaned between each spectra, and the same cuvette was used each time
- Tris buffer serial dilution
- .1M solution of Tris was needed to begin serial dilutions.
- .1M Tris * .1L = .01mol
- .01mol Tris * 121.14g/1mol = 1.21g Tris
- Stock solution of Tris was made at pH 8 and 10.
- 1.21g Tris was added to each tube, with 85mL H2O
- HCl was added dropwise until the pH changed to 8 and 10 in each tube, as indicated by a pH meter. 9mL HCl was added for pH=8 and 2mL was used for ph=10.
- Sufficient water was added to leave the final volume of the solution at 100mL.
- 1mL was taken from the tube with a pipette and moved to the next tube. 1mL from this tube was moved to the next tube, and so on until all get some amount of Tris.
- 9mL H2O was put in the first tube to receive 1mL Tris from the stock solution.
- A serial dilution was repeated with H2O rather than Tris.
- Test tubes were wrapped and heated at at 80°C for 4 hours.
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