User:Daniel-Mario Larco/Notebook/AU Biodesign Lab - 09/03/2013/2014/02/11

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Procedures

  • Remake dopamine coated AuNPs
  1. Dissolve dopamine in 100mL Tris Buffer to have a 10mM solution at pH=8.5
  2. Mix 17mL of AuNP solution with 17mL of the dopamine solution
  3. Stir under air at room temperature for 1 hour
  4. Purify by centrifuging and then redisperse in water.
  5. Filtrate through syringe filters with cellulose acetate membranes with pore size of 1um
  6. Make 2 solutions.
    1. to be stored over night on lab bench
    2. to be stored over night in the refrigerator
  • Testing Activity of Alpha-Method HRP:AuNPs
  1. A solution of H2O2 was prepared by mixing 113uL of 30% H2O2 with 887uL H20
  2. A solution of OPD was prepared by dissolving OPD in H20 to have a final concentration of 1mg/mL
    1. 10mg in 10mL
  3. In order to test the activity of the HRP:AuNP, the HRP:AuNPs were mixed with OPD and H2O2
    1. 0.75mL of H2O2, 2.025mL OPD and 0.225mL of sample
- The UV-Vis spectra showed that in the presence of the HRP:AuNP the OPD peak decreased and another peak appeared. Compared to a sample of OPD alone with H2O2, the sample with the nano particles showed the peak forming at a much faster rate.
- Because the solution may contain some free HRP molecules, we will centrifuge the solution to precipitate the nano particles coated with HRP and test them for activity.



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