- Remake the dopamine addition from 02/05
- Test the activity of citrate-HRP-GNPs using O2 binding assays
Activity of citrate-HRP-GNPs
- We are going to use an assay provided by Dr. Hartings
- The assay uses a reagent that changes color when HRP is added in addition to hydrogen peroxide
- The hydrogen peroxide alone will change the color of the solution, so we will be comparing hydrogen peroxide alone with hydrogen peroxide and GNPs
- We also are going to run UV Vis of the samples to see how the absorption spectra changes over time
- Dissolve dopamine in 100mL Tris Buffer to have a 10mM solution at pH=8.5
- Mix 17mL of GNP solution with 17mL of the dopamine solution
- Stir under air at room temperature for 1 hour
- Purify by centrifuging and then redisperse in water.
- Filtrate through syringe filters with cellulose acetate membranes with pore size of 1um
- Make 2 solutions.
- to be stored over night on lab bench
- to be stored over night in the refrigerator
- The GNPs with HRP were functional
- In the 10^-11 dilution, we observed a color change from deep red initially to a much lighter red
- In our calculations we decided to use the 10^-11 dilution and water as our blank, so we could subtract protein from water.
Protein, OBD, H202
- For each sample run in UV-Vis there were:
- 0.225mL HRP
- 2.25mL OBD stock
- 0.75mL hydrogen peroxide stock