Why is the expression and purification of proteins so import_Protein Technology Column_Technical FAQs_Information Center

A protein must be purified before its structure and the mechanism of its action can be studied. However, because proteins vary in size, charge, and water solubility, no single method can be used to isolate all proteins. To isolate one particular protein from the estimated 10,000 different proteins in a cell is a daunting task that requires methods both for separating proteins and for detecting the presence of specific proteins.

Any molecule, whether protein, carbohydrate, or nucleic acid, can be separated from other molecules based on large differences in some physical characteristic. Although the sequence of amino acids in a protein uniquely determines its function, the most useful physical characteristic for separation of proteins is size, defined as either length or mass. In this section, we briefly outline different techniques for separating proteins based on their size and other properties. These techniques also apply to the separation of nucleic acids and other biomolecules. We then consider general methods for detecting, or assaying, specific proteins, including the use of radioactive compounds for tracking biological activity. Finally, we discuss several techniques for characterizing a protein’s mass, sequence, and three-dimensional structure.

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Why is the expression and purification of proteins so import

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