User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/02: Difference between revisions
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* The required weight for 4-iodophenol is 0.810 g to make 0.0025 M. The actual weighed amount was 0.8109 g. This amount is dissolved in 700 μM dimethyl sulfoxide (DMSO, polar aprotic solvent) due to its immiscibility in water. | * The required weight for 4-iodophenol is 0.810 g to make 0.0025 M. The actual weighed amount was 0.8109 g. This amount is dissolved in 700 μM dimethyl sulfoxide (DMSO, polar aprotic solvent) due to its immiscibility in water. | ||
* Dissolved 25 mg of 4-aminoantipyrine (AAP) into 50 mL of water. | |||
* Weighed out 1 mg of the brown granular solid of horseradish peroxidase (HRP) and dissolved it in 1 mL water. | * Weighed out 1 mg of the brown granular solid of horseradish peroxidase (HRP) and dissolved it in 1 mL water. | ||
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* To obtain 0.0017 M of hydrogen peroxide, added 1 mL 30% of the clear and colorless liquid into 100 mL of water. Then, collected 1 mL from the solution to dilute it into 50 mL of the 0.2 M phosphate buffer. | * To obtain 0.0017 M of hydrogen peroxide, added 1 mL 30% of the clear and colorless liquid into 100 mL of water. Then, collected 1 mL from the solution to dilute it into 50 mL of the 0.2 M phosphate buffer. | ||
* In reference to the document of [http://openwetware.org/wiki/Image:HRP_Assays.docx Novy, Patel, and Wang] | * Placed the settings of the spectrophotometer on kinetics, at 25°C, 510 nm from the methods function. | ||
* In reference to the document of [http://openwetware.org/wiki/Image:HRP_Assays.docx Novy, Patel, and Wang], the group has chosen to prepare the sample solution with the most linearity. The sample solution has the following concentration of reactants. | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'' '' | |||
| align="center" style="background:#f0f0f0;"|''4-iodophenol'' | |||
| align="center" style="background:#f0f0f0;"|''AAP'' | |||
| align="center" style="background:#f0f0f0;"|''H<sub>2</sub>O<sub>2</sub>'' | |||
| align="center" style="background:#f0f0f0;"|''HRP'' | |||
|- | |||
| Molarity||18 mM||156.25 μM||1.7 mM||2.3 μM | |||
|- | |||
| Volume||10 μL|| ||750 μL||50 μL | |||
|- | |||
Revision as of 21:43, 7 October 2012
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Preparation of the Reactants of HRP Oxidase Assay
0.020 mol of Na2HPO4 × [math]\displaystyle{ \frac{268.07g}{1mol} }[/math] of Na2HPO4 = 53.61 g of Na2HPO4
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