User:Floriane Briere/Notebook/CHEM-496/2011/09/14
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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). According to this curve, the 1/1000 diluted MPB solution has a concentration of 0.0312nM (0.026*(6/5) = 0.0312nM). So, the pure MPB solution has a concentration of 31.2nM (0.0312*1000 = 31.2nM).
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) NotesThis area is for any observations or conclusions that you would like to note.
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