Question 4 2 p In lab, you diluted your unknown protein solution by combining 53

Question

Question 4 2 p In lab, you diluted your unknown protein solution by combining 53 iL of unknown protein solution with enough PBS buffer to obtain a total volume of 100 1, then added 1000 IL Bradford reagent. After waiting ten minutes, the absorbance at 595 nm was 0.373. Using the standard curve from the previo question, calculate the total concentration of protein (in 8) in the cuvette. mL Slope 0.02371 Intercept 0.01132 2 0.973 Note: These values are made up and will differ from those you obtain in lab. Remember that your unknown must fall within the range of your standard curve. Addendum: Keep in mind that this is calculating the concentration of protein in the cuvette, which is not the same as the total protein concentration in the original sample.

Explanation / Answer

Question 3

You have information for straight line relationship of absorbance vs concentration the equation of the line is

y = mx + b

where y is the absorbance, m is the slope which is the multiplication of molar absortivity and the pathlenght of the cuvette, b is the intercept

I am using the graph in there and I will only use 2 point

the initial one, x =0 and y = 0

second point, x = 10 and y = 0.15

calculate slope

slope = (y2-y1) / (x2-x1) = (0.15 - 0 ) / (10 - 0) = 0.15 / 10 = 0.015

since the line starts at 0 then

y = 0.015 x where y is absorbance the slope is molarabsortivity * pathlenght of the cuvette

x is the concentration

0.418 = 0.015C

concentration = 0.418 / 0.015 = 27.866 ug / mL

Question 4 is solved the same way if you have different results for slope or the intercept you should use those and apply the same method from question 3

If you solve the question 4 with the numbers I got from the graph then

0.373 = 0.015 * x

x = 0.373 / 0.015 = 24.866 uM / mL

*if this answer is helpful dont forget to rate it =), feel free to comment if you have any doubt

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