User:Floriane Briere/Notebook/CHEM-496/2011/09/14: Difference between revisions
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And we know that c = 0,0312nM and l = 1cm | And we know that c = 0,0312nM and l = 1cm | ||
This curve is made with the 1/1000 diluted MPB solution measures. It shows MPB molar absorbitivity as a function of the wavelength. | |||
==Notes== | ==Notes== |
Revision as of 09:05, 20 September 2011
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ObjectiveToday's experiment is to test the Bradford Assay technique which is going to allow us to determine the unknown proteins’ concentration of a solution. We are going to use 6 standards solutions of BSA whose concentrations are known. Then, thanks to a spectrophotometer, we’ll be able to determine the concentration of the MBP solution. Protocol
Results* Curve of the concentration (nM) as a function of the Absorbance at 595nm Red lines represent the Absorbance at 595nm of the 1/1000 diluted MPB solution. This curve allow us to determine the concentration of the 1/1000 diluted MPB solution. To draw this curve, we add to take into account that the Bradford Assay was made with 5/6 diluted solutions (because we added 1ml of solution to 200µl of Bradford reagent). We can also calculate the unknown concentration thanks to the line’s equation (y = 2.5734x + 0.3878): Abs(595nm) = 2.5734 * concentration + 0.3878 => concentration = (0.44 - 0.3878)/2.5734 = 0.0202nM According to this curve, the 1/1000 diluted MPB solution has a concentration of 0.0243nM (0.0202*(6/5) = 0.0243nM). So, the pure MPB solution has a concentration of 24.3nM (0.0243*1000 = 24.3nM).
This curve is made with the 1/1000 diluted MPB solution measures.
According to Beer-Lambert law, Absorbance = molar absorptivity (L.mol^-1.cm^-1) * concentration (M) * length of the cuve (cm) So, molar absorptivity (ε) = Absorbance/(concentration (c)* length of the cuve (l)) And we know that c = 0,0312nM and l = 1cm This curve is made with the 1/1000 diluted MPB solution measures. It shows MPB molar absorbitivity as a function of the wavelength. NotesThis area is for any observations or conclusions that you would like to note.
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