Use the following available information answer questions 1-5. Protein name Molecular mass pl Subunit structure Protein A 250,000 9.7 Monomer Protein B 75,000 13,000 46,000 4.8 Tetramer Protein C 3.1 Monomer Protein D 10.2 Monomer 1. Name one purification method that can separate Protein A from a solution that contains all 4 proteins. 2. If you do ion-exchange chromatography at pH 7 on a column of diethylaminoethyl (DEAE)-cellulose, which of the above proteins will bind to the column? 3. If you do ion-exchange chromatography at pH 7 on a column of carboxymethyl (CM)-cellulose, which of the above proteins will not bind to the column? 4. If you do isoelectric focusing on a mixture of the above proteins followed by SDS-PAGE, and stain the gel with Coomassie Blue, how many protein spots do you expect to see? Explain your answer. 5. If I saw 5 spots in the experiment described in question 4, how will you explain the results?
Proteins
We generally tend to think of proteins only from a dietary lens, as a component of what we eat. However, they are among the most important and abundant organic macromolecules in the human body, with diverse structures and functions. Every cell contains thousands and thousands of proteins, each with specific functions. Some help in the formation of cellular membrane or walls, some help the cell to move, others act as messages or signals and flow seamlessly from one cell to another, carrying information.
Protein Expression
The method by which living organisms synthesize proteins and further modify and regulate them is called protein expression. Protein expression plays a significant role in several types of research and is highly utilized in molecular biology, biochemistry, and protein research laboratories.
Name one purification method that can separate Protein A from a solution that contains all four proteins. (Size exclusion chromatography, ion exchange chromatography, Purification by affinity chromatography, High pressure liquid chromatography, Isoelectric focusing, SDS-PAGE, or Two-dimensional Electrophoresis)
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