User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/02: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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* Placed the settings of the spectrophotometer on kinetics, at 25°C, 510 nm from the methods function. | * 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 (MDK)], the group has chosen to prepare the sample solution with the most linearity. The sample solution has the following concentration of reactants listed in the table below. | * In reference to the document of [http://openwetware.org/wiki/Image:HRP_Assays.docx Novy, Patel, and Wang (MDK)], the group has chosen to prepare the sample solution with the most linearity. The sample solution has the following concentration of reactants listed in the table below. The original concentration of AAP was 2.5 mM amounting to 700 μL. | ||
* Transferred the reactants to a plastic disposable sample cell using a micropipet. NOTE: The HRP was added last when the sample was already inside the chamber. The addition of HRP initiates the reaction. | * Transferred the reactants to a plastic disposable sample cell using a micropipet. NOTE: The HRP was added last when the sample was already inside the chamber. The addition of HRP initiates the reaction. | ||
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* For the following day, the agenda would be to ascertain the appropriate volume of AAP while keeping the same concentration. | * For the following day, the agenda would be to ascertain the appropriate volume of AAP while keeping the same concentration. | ||
{| {{table}} | {| {{table}} | ||
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==HRP Assay Figures== | ==HRP Assay Figures== | ||
* Charts embedded below represent the HRP Assay runs that have varying concentrations of HRP and AAP. The variations in the physical condition of the surrounding environment were done to investigate the appropriate ratio between HRP and AAP. | |||
[[Image:HRP10022012.png]] | |||
* Runs 1, 3, and 4 reflect absorbance trends where the reaction had a low concentration of HRP. | |||
* Run 2 shows the reaction where there was an error in calculating the proper volume for the HRP and AAP ratio. The group added 50 μL of HRP against a very low concentration of AAP. | |||
* Run 5 is the replicated absorbance from MDK using the standard condition where the volume of AAP is 700 μL and HRP is 50 μL. | |||
[[Image:DviewHRP10022012.png]] | |||
* The rates of runs 2 and 5 are shown below to indicate that the rate of run 5 (reaction with the proper amount of HRP) has a linear rate compared to run 2. | |||
[[Image:Ratesofrxnrun2&5.png]] | |||
[[Image:Dviewratesofrxnrun2&5.png]] | |||
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Latest revision as of 22:06, 26 September 2017
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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
HRP Assay Figures
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