The amount of protein in a sample can be quantified by several kinds of spectrop

Question

The amount of protein in a sample can be quantified by several kinds of spectrophotometric methods. One method is based on the amount of light directly absorbed by the protein (no added reagents), while other methods utilize a chromophore-containing reagent and measure light absorbed by protein-reagent complexes. Match the descriptions with the appropriate method.

Match the descriptions with the appropriate method.

Methods:

Measures protein absorbance directly

Measures absorbance of protein reagent complexes

Applies to both methods

Descriptions:

Detection wavelenth of 280 nm, the max absorbance for aromatic side chains

Absorbance intensity is highly dependent on presence of Trp and Tyr residues

Absorbance intensity is independent of protein amino acid composition
Detection wavelength set at max absorbance for bound dye

Nucleic acids absorb light at detection wavelength so correction factor must be applied

Sample concentration is determined by comparison to standard curve generated with a pure protein

Absorbance is proportional to solute concentration for a fixed path length

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Explanation / Answer

Measures protein absorbance directly -  Absorbance intensity is highly dependent on presence of Trp and Tyr residues

Measures absorbance of protein reagent complexes - Detection wavelength set at max absorbance for bound dye

Nucleic acids absorb light at detection wavelength so correction factor must be applied

Applies to both methods -  Absorbance is proportional to solute concentration for a fixed path length

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