Results table Bradford (mL) BSA (μL) Absorbance 3 0 0 3 10 0.228 3 20 0.402...

Results table

Bradford (mL)	BSA (μL)	Absorbance
3	0	0
3	10	0.228
3	20	0.402
3	30	0.553
3	40	0.715
3	50	0.776
3	30	0.58 unknown

Graph your standard curve by plotting protein content in the abscissa and absorbance in the ordinate.
Calculate the protein content of your samples.

Expert Solution

The Bradford assay is based on an absorbance change of the dye Coomassie Brilliant Blue. The more protein present, the more Coomassie binds. The Bovine Serum Albumin (BSA) is the most widely used protein as the standard to calculate the protein content in a sample.

To know the protein content we must plot the protein content and the absorbance:

We must calculate a linear regression curve through these points to determine the equation of the straight line in the form of "y = mx + b" where y = absorbance and x = protein concentration. Several scientific graphing packages calculate this equation, in this case, I used excel to obtain the equation, which is:

y = 0.0157x + 0.0534

With a coefficient of determination R² ≥ 0.95, that means that is a reliable linear regression curve

Then, we can calculate the protein content by knowing the "y" value which is 0.58 (the absorbance value)

We clear the "x" from the equation and we get

x = (y - 0.0534) / 0.0157

Substitute "y" value

x = ( 0.58 - 0.0534) / 0.0157

x = 33.54

The absorbance from the unknown sample was 0.58, then, its protein content is 33.54

gladiator answered 2 hours ago

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