User:Eleni N. Kalivas/Notebook/CHEM-571/2013/10/16

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Objective

To test the activity of HRP-NPs for the catalytic conversion of luminol. We would like to ascertain what percent of HRP is still active.

Protocol

The following stock solution information was taken from Matt Harting's notebook.


Stock Solution

  1. Buffer
    1. 0.6140g Tris in 1L, pH set to 8 with HCl ---> 5.1mM
  2. Luminol
    1. Dissolve 12.9mg luminol in 300uL of DMSO
    2. Add to 50mL buffer ---> 1.46mM
  3. H2O2
    1. 312uL 30% H2O2 into 100mL buffer ---> Should be 45mM
    2. Check absorption at 250 source (ε(250) = 16.69 M-1cm-1). A = 0.7392. [H2O2] = 44.29mM

Protocol Notes:

  1. The final concentration of luminol is such that the absorbance at 333nm is less than one
  2. The final concentration of hydrogen peroxide is fifty times that of the luminol.
  3. Using the 60:1 [Au:HRP] solution, 100uL will be used
  4. Add buffer so that the final concentration is 3mL

Data

  • The Absorbance of BSA-AuNPs could not be done because the material crashed out of the solution. Therefore new BSA-AuNP solution is being made, and will be placed into the oven for four hours, to be tested tomorrow.
  • Absorbance Spectra of Citrate-AuNPs and HRP-AuNPs

  • Figure 1. Absorbance spectra of the catalytic activity of HRP-AuNPs



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