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(New page: ==Objective== Synthesize two different sets of gold nanoparticles. In one set, Au3+ is reduced by a protein (bovine serum albumin, BSA) and the synthesized nanoparticle is also surrounded...)
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Synthesize two different sets of gold nanoparticles. In one set, Au3+ is reduced by a protein (bovine serum albumin, BSA) and the synthesized nanoparticle is also surrounded and stabilized by BSA. In the second set, Au3+ is reduced by citrate, and the AuNP is stabilized by citrate in solution. The BSA-AuNPs are purple in color and the citrate-AuNPs are more of a burgundy (reddish) color.
- (1) 10 ml volumetric flask
- (1) 50 mL volumetric flask
- 2.54mM gold (HAuCl4)
- 0.0156mM BSA
- deionized water
- aluminum foil
- plastic falcon test tube
- hot plate
- 1% (w/v) sodium citrate
Start by placing all of your materials into a volumetric flask so that you know the exact volumes and exact amounts of what you have added. I will have prepared the initial protein and gold solutions for you (this one time only). In the future, you will be expected to make most of the starting solutions. This procedure was taken from the following reference and has been used by our previous two Experimental Biological Chemistry groups.
- Add 1.2mL of the (~2.54mM -note the exact concentration) gold (HAuCl4) solution to a 10mL volumetric flask
- Add an appropriate amount of BSA solution so that the final concentration of gold is 90X that of BSA
- Add deionized water up to 10.2mL
- Transfer solution to a test tube and cap with aluminum foil
- Heat in oven at 80C for 3 hours
- Transfer solution to a plastic falcon tube (with blue cap)
Stock solutions made
- Gold solution (HAuCl4·3H2O) 0.0100g in 0.0100mL water → 2.54mM
- BSA solution 0.0104g BSA (MW = 66776g/mol) in 0.0100mL water → 15.6μM
This procedure is used by Dr. Miller in her research lab - Allison Alix's notebook and is taken from this reference and can be analyzed according to this reference.
- Take 50mL of the HAuCl4 solution from the 250mL volumetric flask. Note the concentration
- Heat this solution to boiling while stirring
- Add 3mL 1.5mL of 1% (w/v) sodium citrate
- Boil solution for another 40 minutes
- Cool to room temperature and measure the volume
- Determine the final concentration of gold and citrate
Note: According to Allison, and Madeline will corroborate, we don't need to reflux the solution (which would carry out the reaction in a closed system). We should be able to bring the solution to a boil on the bench top.
Stock solutions made
- Gold solution (HAuCl4·3H2O) 0.0245g in 0.2505mL water → 0.249mM
- Sodium Citrate (Na3C6H5O7·2H2O) 0.1010g in 10.0mL → 1.01%
- Who has experience with this protocol?
or instead, discuss this protocol.