User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/07: Difference between revisions
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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]]. | ||
*Prepare Au/HRP solutions | |||
==Procedure== | ==Procedure== | ||
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* The absorbance values at 595 nm of each solution were graphed versus BSA concentration in mg/mL to produce a calibration curve. | * The absorbance values at 595 nm of each solution were graphed versus BSA concentration in mg/mL to produce a calibration curve. | ||
*Note that : When these samples were originally run, they produced absorbances over 1 which resulted in us needing to dilute the solution further. As a result, all solutions were diluted by a half to 715μL of original solution with an additional 715μL of water. | |||
* After the calibration curve was made for the standards, the concentration of the ADA collected on the two days were analyzed. We ran both fracs that possessed the majority of the sample from [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26| 2012/09/26]] and only frac number 2 from [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/06| 2012/11/06]] | |||
* 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) | |||
* For all information regarding 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 | |||
|} | |||
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Revision as of 11:03, 15 November 2012
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Bradford AssayGoals
Procedure
DATA
Concentrations, absorbances, for the collected fracs of ADA
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