Objective
- Obtain UV/Vis spectra for HRP assay with absorbance at 510nm plotted against time
Procedure
- Concentrations will be kept constant from HRP assays done by another group in |AU's Biomaterial Design Lab with nothing changed except for the addition of varying concentrations of gold nanoparticles. Their writeup can be found here: File:HRP Assays.pdf. The concentration of HRP was raised by 150% because it did not produce a peak at first.
- For six cuvettes to have 1.7mM hydrogen peroxide, 18μM iodophenol, 156.25μM AAP and 2.3μM HRP, the following volumes of the respective stock solutions (made 10/02) were added:
- 750μL 30% hydrogen peroxide solution
- 10μL .17M iodophenol solution
- 35μL 2.5mM AAP solution
- 22μL 23μM HRP solution
Amount of gold and water used in each cuvette
| AuNP solution (μL) |
water (μL)
|
| 0 |
630
|
| 20 |
610
|
| 40 |
590
|
| 60 |
570
|
| 80 |
550
|
| 100 |
530
|
- The amount of stock solution needed of each reagant was calculated as such:
- M1V1=M2V2
- .023mM HRP stock * .001L = .0023mM HRP * volume needed
- 22μL HRP stock needed
Data
We are waiting for the spectra to be uploaded by the researcher who has it. It will be posted here once available, and reaction rates will be calculated.
- Abigail E. Miller 11:49, 7 October 2012 (EDT):why don't you have it? it is not an excuse for it not being in your notebook.
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