User:Helen L. Slucher/Notebook/CHEM-572/2014/02/11

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Objective Remake the dopamine addition from 02/05 Test the activity of citrate-HRP-GNPs using O2 binding assays Procedure 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 Dopamine Coating 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 Data 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