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Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer

Published on: November 4, 2009

Spectrophotometric methods to determine protein concentration.

J P Dean Goldring¹

¹Biochemistry, University of KwaZulu-Natal, Pietermaritzburg, PBX01, Scottsville, 3209, South Africa, GoldringD@ukzn.ac.za.

Methods in Molecular Biology (Clifton, N.J.)

|June 6, 2015

Summary

Accurate protein concentration measurement is vital for enzyme studies and protein isolation. This review covers spectrophotometry and dye-binding methods for determining protein concentration in solution.

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Area of Science:

Biochemistry
Analytical Chemistry

Background:

Protein concentration is crucial for enzyme activity assays and monitoring protein purification.
Assessing isolation procedure efficacy requires understanding specific protein levels relative to total protein content.

Purpose of the Study:

To review common methods for determining protein concentration in solution.
To provide an overview of spectrophotometric and dye-binding techniques.

Main Methods:

Spectrophotometry in the ultraviolet (UV) range.
Colorimetric assays involving dyes that interact with proteins.
Methods utilizing copper-protein interactions.

Main Results:

Spectrophotometry offers a direct measure of protein concentration.
Dye-binding and copper-based methods provide indirect quantification through color changes.
Each method has specific advantages and limitations regarding sensitivity, interference, and cost.

Conclusions:

Various protein determination methods exist, each suited for different experimental contexts.
Choosing the appropriate method depends on factors like sample purity, required sensitivity, and available equipment.
Accurate protein quantification is fundamental for reliable biochemical and biophysical analyses.