User:Michael F. Nagle/Notebook/Chem 571/2012/09/05: Difference between revisions

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###15μM*(volume [[AU_Biomaterials_Design_Lab:Materials/BSA|BSA]] stock solution) = (1.5*6)
###15μM*(volume [[AU_Biomaterials_Design_Lab:Materials/BSA|BSA]] stock solution) = (1.5*6)
####0.6mL [[AU_Biomaterials_Design_Lab:Materials/BSA|BSA]] stock solution in each tube
####0.6mL [[AU_Biomaterials_Design_Lab:Materials/BSA|BSA]] stock solution in each tube
###7520μM*x mL=6mL*(1.5*60μM)
####0.069mL [[AU_Biomaterials_Design_Lab:Materials/HAuCl4|HAuCl<sub>4</sub>]] stock solution
####5.331 mL H<sub>2</sub>O
###7520μM*x mL=6mL*(1.5*120μM)
###7520μM*x mL=6mL*(1.5*120μM)
####0.143mL [[AU_Biomaterials_Design_Lab:Materials/HAuCl4|HAuCl<sub>4</sub>]] stock solution
####0.143mL [[AU_Biomaterials_Design_Lab:Materials/HAuCl4|HAuCl<sub>4</sub>]] stock solution

Revision as of 11:21, 11 September 2012

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Objectives

  • Using UV/Vis, find which mole ratios of HAuCl4 to BSA result in the most BSA unfolding and formation of gold nanoparticles.
  • Make solutions with varying concentrations of Tris buffer and HAuCl4 and BSA fibers

Procedure

  1. UV/Vis
    1. 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
    2. Spectra were obtained and peaks for gold nanoparticles were identified at the 520nm range.
    3. Cuvette was cleaned between each spectra, and the same cuvette was used each time
  2. Tris buffer serial dilution
    2. A partner did the serial dilution while I was doing UV/Vis and I appear to have copied a calculation down wrong because the henderson-hasselbach equation does not add up.
    3. 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.
    4. 9mL H2O was put in the first tube to receive 1mL Tris from the stock solution.
    5. A serial dilution was repeated with H2O rather than Tris.
    6. Test tubes were wrapped and heated at at 80°C for 4 hours.
  3. Au/BSA varying mole ratios
    1. A stock solution of HAuCl4 was made, with the attempted Molarity of 10mM.
      1. (moles HAuCl4)/.025L H2O = .01M
      2. 2.5*10-4 mol HAuCl4
      3. 2.5*10-4 mol * 339.785mol/g HAuCl4 = .0849g
      4. .0283g of the .0924g stuck to the weigh paper, leaving .0639gHAuCl4.
      5. .0639g HAuCl4 * (1molHAuCl4/339.79g/molHAuCl4)= xM HAuCl4
      6. .000188mol/.025L=xM HAuCl4
      7. .007520M HAuCl4
      8. .0639g HAuCl4 was put in 25mL to make a 10mM solution
    2. A 15μM stock solution of BSA was made
      1. (moles BSA)/.025L = .000015
      2. 3.75*10^-7mol BSA
      3. 3.75*10^-7mol BSA * 66,463g/mol BSA = .0249g BSA
      4. .0249g BSA was put in 25mL to make a 15μM solution
    3. Solutions were made with the mole ratios of (HAuCl4/BSA) 120, 128, 130, 132, 133, and 134. Volume of HAuCl4 stock solution was calculated for each tube and inserted. The water needed for each tube was calculated by subtracting the volume of HAuCl4 stock solution and BSA stock solution from 6mL.
      1. m1v1=m2v2
      2. 15μM*(volume BSA stock solution) = (1.5*6)
        1. 0.6mL BSA stock solution in each tube
      3. 7520μM*x mL=6mL*(1.5*120μM)
        1. 0.143mL HAuCl4 stock solution
        2. 5.257mL H2O
      4. 7520μM*x mL=6mL*(1.5*128μM)
        1. 0.153mL HAuCl4 stock solution
        2. 5.247mL H2O
      5. 7520μM*x mL=6mL*(1.5*130μM)
        1. .155mL HAuCl4 stock solution
        2. 5.245mL H2O
      6. 7520μM*x mL=6mL*(1.5*132μM)
        1. 0.157mL HAuCl4 stock solution
        2. 5.243mL H2O
      7. 7520μM*x mL=6mL*(1.5*133μM)
        1. 0.159mL HAuCl4 stock solution
        2. 5.241mL H2O
      8. 7520μM*x mL=6mL*(1.5*134μM)
        1. 0.160mL HAuCl4 stock solution
        2. 5.240mL H2O
  1. The tubes were wrapped in tin foil and heated at 80°C for 4 hours.


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