User:Floriane Briere/Notebook/CHEM-496/2012/04/04: Difference between revisions
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5ml of water + 80µL of DMSO => Total volume = V = 0.0058L | 5ml of water + 80µL of DMSO => Total volume = V = 0.0058L | ||
MW of the dye = 573.51 g/mol | MW of the dye = 573.51 g/mol | ||
So, n = 10^-3/573.51 = 1.74* 10^-6 moles | So, n = 10^-3/573.51 = 1.74* 10^-6 moles | ||
And, C = (1.74* 10^-6)/0.0058 = 3*10^-4 M | And, C = (1.74* 10^-6)/0.0058 = 3*10^-4 M | ||
# Determination of the molar absorptivity of the dye using Beer's law. | # Determination of the molar absorptivity of the dye using Beer's law. | ||
We are going to use the Absorbance at 602nm (which is the absorption value given by the dye manufacturer); at this wavelength, fluctuation should be the lowest. | We are going to use the Absorbance at 602nm (which is the absorption value given by the dye manufacturer); at this wavelength, fluctuation should be the lowest. |
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ObjectiveProtocol
DataOne of our objective is to determine the number of Lysine (on the BSA molecule) which are bound to a dye molecule. To do so, we need to evaluate the concentration of dye in our solution and the amount of BSA protein. The amount of dye in each solution can be determined using UV spectrum; the dye in solution has a known-concentration which will allow us to find the molar absorptivity of the dye (at 602nm and 625nm; Gold particles also absorb around 600nm and we want to lower this effect as much as possible).
The dye in solution has been prepared on the 21/2 using: m = 1mg of dye 5ml of water + 80µL of DMSO => Total volume = V = 0.0058L MW of the dye = 573.51 g/mol
We are going to use the Absorbance at 602nm (which is the absorption value given by the dye manufacturer); at this wavelength, fluctuation should be the lowest. Molar absorbtivities (ε, L/mol/cm) were determined using Beer's law: A = ε*l*C With A = absorbance at 602nm l = width of the cuvette (1cm) C = dye concentration (3*10^-4 M) The molar absorptivity of our dye at 602nm is equal to = 3419 L.mol^-1.cm^-1 (It's determined by doing the average of the three different ε values)
Molar absorbtivities (ε, L/mol/cm) were determined using Beer's law: A = ε*l*C With A = absorbance at 625nm l = width of the cuvette (1cm) C = 3*10^-4 M [[ NotesThis area is for any observations or conclusions that you would like to note.
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