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.
The following stock solution information was taken from Matt Harting's notebook.
- 0.6140g Tris in 1L, pH set to 8 with HCl ---> 5.1mM
- Dissolve 12.9mg luminol in 300uL of DMSO
- Add to 50mL buffer ---> 1.46mM
- 312uL 30% H2O2 into 100mL buffer ---> Should be 45mM
- Check absorption at 250 source (ε(250) = 16.69 M-1cm-1). A = 0.7392. [H2O2] = 44.29mM
- The final concentration of luminol is such that the absorbance at 333nm is less than one
- The final concentration of hydrogen peroxide is fifty times that of the luminol.
- Using the 60:1 [Au:HRP] solution, 100uL will be used
- Add buffer so that the final concentration is 3mL
- 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