User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/03

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As mentioned in the previous entry of Oct. 2, there was difficulty in finding the appropriate ratio between AAP and HRP.
Using the Kinetics program, trial and error was done to find a linear trend for the absorbance of HRP. The calculated volume was 22 μL. The optimal volume for HRP was found at 33 μL. This can be seen on the graph below.

Now that the optimal amount of HRP was found, the reaction of AAP and hydrogen peroxide with HRP was tested if this was affected by the addition of Au/Citrate. The first run is HRP alone with the reaction followed by the addition of Au/Citrate as shown below.
Au/Citrate was made from 20 mL of 1 mM HAuCl₄ with 20 mL of 1% citrate. The Au/Citrate was added in multiple of 20. Each 20 μL is equivalent to 2%.

Trials	'	'	'	'	'	'
Cuvet	hydrogen peroxide concentration (mM)	Iodophenol in DMSO concentration (μM)	AAP Concentration (μM)	HRP concentration (μM)	μL of gold np added	μL of water added
1( standard)	1.7	18	156.25	3.45	0	630
1	1.7	18	156.25	3.45	20	610
2	1.7	18	156.25	3.45	40	590
3	1.7	18	156.25	3.45	60	570
4	1.7	18	156.25	3.45	80	550
5	1.7	18	156.25	3.45	100	530
Total volume added	750 μL	10 μL	35 μL	33 μL

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-* Now that the optimal amount of HRP was found, the reaction of AAP and hydrogen peroxide with HRP was tested if this was affected by the addition of Au/Citrate. The first run is HRP alone with the reaction followed by the addition of Au/Citrate.
+* Now that the optimal amount of HRP was found, the reaction of AAP and hydrogen peroxide with HRP was tested if this was affected by the addition of Au/Citrate. The first run is HRP alone with the reaction followed by the addition of Au/Citrate as shown below.
+* Au/Citrate was made from 20 mL of 1 mM HAuCl<sub>4</sub> with 20 mL of 1% citrate. The Au/Citrate was added in multiple of 20. Each 20 μL is equivalent to 2%.