Data Analysis 1. Convert your absorbances, A, of ferritin and cytochrome c into
ID: 551561 • Letter: D
Question
Data Analysis 1. Convert your absorbances, A, of ferritin and cytochrome c into concentrations using the following extinction (mg/mL) for both ion-exchange and gel filtration coefficients, , and Beer's Law where A- ebc. The path length, b, is 1 cm and c is the concentration in mg/mL. Report your data in tables. NOTE: It cannot assumed that the sample run from any column is pure ferritin or cytochrome c Fortunately, absorbances are additive and any absorbance reading can be assumed to come from both ferritin and cytochrome c. For each run, two equations and two unknowns must be used to find the concentration of each protein. Be sure to include sample calculations. be | at 410nm (in mL/mg*am) l e at 340nm (in mL/mg*am) Ferritin 1.107 2.551 Cytochrome c 8.005 1.747 For Example: Given that the Absorbance for GF sample fraction number 3 at 410 nm is O.76 and at 340 nm is 0.21, find the concentration of the proteins. The path length of the cuvette is 1cm and extinction coefficients are given above. Let x "concentration" of ferritin in mg/mL Let y = "concentration" of cytochrome c in mg/mL Atotal = Ebc (ferritin) + ebc (cytochrome c) A 410 nm = 0.76 = 1.107 x + 8.005 y and A 340 nm = 0.21 = 2.551 x + 1.747 y Doing the math... x = 0.0191 mg/mL and y = 0.0923 mg/mL NOTE: In the interest of saving time this system of equations can be solved quickly by setting up a spreadsheet equationExplanation / Answer
To do in excel, you will Need Solver Add-in ( which can be found in Data Tab):
Choose three blocks A, B and C to work with.
Now put your equations in A block as-
=(1.107*C1)+(8.005*C2) in A1
=(2.551*C1)+(1.747*C2) in A2
Now put B1 = 0.76 and B2 = 0.21
After that open Solver tab:
In By Changing Variable Cell put: Select blocks C1:C2
Then Add subject to constraints by putting-
$A$1 = $B$1
$A$2 = $B$2
Then select the Make unconstrained variables non-negative option.
After that click on Solve and OK.
You will get the solutions in C1 and C2 block.
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