biochemistry laboratory student used the Bradford protein assay to measure the l

Question

biochemistry laboratory student used the Bradford protein assay to measure the lysozyme (protein) content in egg whites. The Bradford protein assay is a spectrophotometric technique that uses a dye, Coomassie Brilliant Blue, whose absorption undergoes a spectroscopic shift when bound to a protein. In the unbound state, Coomassie Brilliant Blue displays a red color. As protein binds to the dye, its absorbance at 595 nm increases and the dye changes to a blue color. The student prepared a 3.000 mL sample containing 2 mg/mL Coomassie Brilliant Blue. To this sample, 200.0 mu L of 45.0 mu g/mL lysozyme was added and an absorbance of 0.247 was measured at 595 nm. Calculate the corrected absorbance for this sample. Corrected absorbance =

Explanation / Answer

Apply:

Corrected Absorbance = Total volume / Initial Volume * Observed Absorbance

so

Total V = 3.0 mL + 600 miuL = 3 mL + 0.6 mL = 3.6 mL

Then

Initial V = 3.0 mL

Observed absorbance = 0.247

Substitute data

Corrected Abs = 3.6/3*0.247 = 0.2964

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