User:Daniel-Mario Larco/Notebook/AU Biodesign Lab - 09/03/2013/2014/02/11: Difference between revisions
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== | ==Procedures== | ||
* | *Remake dopamine coated AuNPs | ||
#Dissolve dopamine in 100mL Tris Buffer to have a 10mM solution at pH=8.5 | |||
#Mix 17mL of AuNP 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 | |||
*Testing Activity of Alpha-Method HRP:AuNPs | |||
#A solution of H2O2 was prepared by mixing 113uL of 30% H2O2 with 887uL H20 | |||
#A solution of OPD was prepared by dissolving OPD in H20 to have a final concentration of 1mg/mL | |||
##10mg in 10mL | |||
#In order to test the activity of the HRP:AuNP, the HRP:AuNPs were mixed with OPD and H2O2 | |||
##0.75mL of H2O2, 2.025mL OPD and 0.225mL of sample | |||
- The UV-Vis spectra showed that in the presence of the HRP:AuNP the OPD peak decreased and another peak appeared. Compared to a sample of OPD alone with H2O2, the sample with the nano particles showed the peak forming at a much faster rate. | |||
- Because the solution may contain some free HRP molecules, we will centrifuge the solution to precipitate the nano particles coated with HRP and test them for activity. | |||
Revision as of 16:22, 17 April 2014
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Procedures
- The UV-Vis spectra showed that in the presence of the HRP:AuNP the OPD peak decreased and another peak appeared. Compared to a sample of OPD alone with H2O2, the sample with the nano particles showed the peak forming at a much faster rate. - Because the solution may contain some free HRP molecules, we will centrifuge the solution to precipitate the nano particles coated with HRP and test them for activity.
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