User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/07: 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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==Goals== | ==Goals== | ||
* Conduct a bradford assay on the collected fraction of ADA made on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/06| 2012/11/06]] and on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26| 2012/09/26]]. | * Conduct a bradford assay on the collected fraction of ADA made on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/06| 2012/11/06]] and on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26| 2012/09/26]]. The bradford assay will allow us to calculate the concentration of ADA present in our collected fraction samples. | ||
*Prepare Au/HRP solutions | |||
==Procedure== | ==Procedure== | ||
* A stock solution of 1mg/mL BSA was prepared | * A stock solution of 1mg/mL BSA was prepared (actual concentration = 1.02mg/mL). 10.2mg BSA added to 10mL of water in a volumetric flask. | ||
*In order to determine the concentration of the ADA collected in the fracs, standard solutions had to be made and run through the UV-Vis first. Seven standard solutions were prepared with 1X Bradford Reagent, varying amounts of BSA stock, and autoclaved water. These solutions were analyzed in the UV-Vis from 500-800 nm. | *In order to determine the concentration of the ADA collected in the fracs, standard solutions had to be made and run through the UV-Vis first. Seven standard solutions were prepared with 1X Bradford Reagent, varying amounts of BSA stock, and autoclaved water. These solutions were analyzed in the UV-Vis from 500-800 nm. | ||
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* In a cuvet, 7.5μL of autclaved water, 7.5 μL of a fraction and 715μL 1X bradford Reagent were added. In total 3 cuvet samples were run through the UV-Vis. | * In a cuvet, 7.5μL of autclaved water, 7.5 μL of a fraction and 715μL 1X bradford Reagent were added. In total 3 cuvet samples were run through the UV-Vis. | ||
**Because the spectra for frac #8 and Frac New had an absorbance of over 1, those cuvet samples were diluted in half using the same procedure as for the standard solutions in order to get an acceptable (below 1) absorbance) | **Because the spectra for frac #8 and Frac New had an absorbance of over 1, those cuvet samples were diluted in half using the same procedure as for the standard solutions in order to get an acceptable (below 1) absorbance) | ||
* For all information regarding the making and analyzing of the Au/HRP solutions , please see [[User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/11/07| Dhea's Notebook]] | |||
==DATA== | ==DATA== | ||
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* Linear regression was used to determine the equation of the line | * Linear regression was used to determine the equation of the line | ||
Concentrations, absorbances, for the collected fracs of ADA | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Sample Name''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume of ADA[mL]''' | |||
| align="center" style="background:#f0f0f0;"|'''Absorbance''' | |||
| align="center" style="background:#f0f0f0;"|'''[ADA] [mg/mL]''' | |||
| align="center" style="background:#f0f0f0;"|'''Mass of ADA[mg]''' | |||
| align="center" style="background:#f0f0f0;"|'''Mols of ADA [mol]''' | |||
| align="center" style="background:#f0f0f0;"|'''Molar absorptivity [mL/mg*cm-1]''' | |||
| align="center" style="background:#f0f0f0;"|'''molarity[M]''' | |||
| align="center" style="background:#f0f0f0;"|'''uM''' | |||
|- | |||
| Frac 2||0.015||0.93||0.017747193||0.025644694||6.30091E-10||52.40265306||4.20061E-05||42.0060507 | |||
|- | |||
| Frac 8||0.0075||0.757||0.011481347||0.016590547||4.0763E-10||65.93302839||5.43507E-05||54.35068601 | |||
|- | |||
| Frac new||0.0075||0.819||0.013726911||0.019835386||4.87356E-10||59.66382586||6.49808E-05||64.98078863 | |||
|} | |||
** Frac 2 represents the flasks 2-4 collection done in [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26| September]]. Frac 8 represents collection of [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26|flask 1]], and Frac New represents the ADA just made [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/06| yesterday.]] | |||
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Bradford AssayGoals
Procedure
DATA
Concentrations, absorbances, for the collected fracs of ADA
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