Chapter 6: Techniques of Protein and Nucleic Acid

Purifications
Matching
A.) hydrophobic
B.) chromophore
C.) foaming
D.) high level expression
E.) 2-mercaptoethanol
F.) positive charge
G.) cation exchange
H.) enzyme-linked immunosorbent assay

1. If antibodies to the protein being assayed are available, a(n) _____ can be developed.
Ans: H
Level of Difficulty: Moderate
Section: 6-1. Protein Isolation

2. ______ chromatography is a method of fractionating a protein mixture according to the

different polarities of the proteins.
Ans: A
Level of Difficulty: Moderate
Section: 6-3. Chromatographic Separations

3. In order for DEAE to act as an anion exchanger, it must have a ______.

Ans: F
Level of Difficulty: Moderate
Section: 6-3. Chromatographic Separations

4. In ______ chromatography, a protein mixture must be applied to the column at a low pH so

that the proteins will have a net positive charge and bind to the column.
Ans: G
Level of Difficulty: Moderate
Section: 6-3. Chromatographic Separations
5. In SDS-PAGE electrophoresis, disulfide-linked protein subunits are separated by first reacting
them with ______.
Ans: E
Level of Difficulty: Easy
Section: 6-4. Electrophoresis

Multiple Choice
6. A fast and common method for determining the concentration of protein in aqueous solution
is:
A) tandem mass spectrometry.
B) salting in with ammonium sulfate.
C) drying a portion and weighing the solid.
D) measuring light absorption at 280 nm.
E) Edman degradation.
Ans: D
Level of Difficulty: Easy
Section: 6-2. Solubilities of Proteins

7. The salting in of proteins can be explained by:

A) salt counter-ions reducing electrostatic attractions between protein molecules.
B) proteins attracting primarily salt cations.
C) proteins attracting primarily salt anions.
D) releasing hydrophobic proteins from nonpolar tissue environments.

E) hydration of the salt ions reducing solubility of proteins.

Ans: A
Level of Difficulty: Moderate
Section: 6-2. Solubilities of Proteins

8. A first step in purifying a protein that was initially associated with fatty substances would be:
A) Coomassie Brilliant Blue dye staining.
B) analytical ultracentrifugation.
C) ELISA.
D) Western blotting.
E) hydrophobic chromatography.
Ans: E
Level of Difficulty: Easy
Section: 5.2.C
Learning objective: Protein Purification and Analysis
9. Which of the following assays would be most specific for a unique protein?
A) Bradford assay
B) UV absorptivity
C) radioimmunoassay
D) molar absorptivity
E) amino acid analysis
Ans: C
Level of Difficulty: Easy
Section: 5.1.A
Learning objective: Polypeptide Diversity

10. An enzyme-linked immunosorbent assay requires:

A) a radioactive substrate.

B) a coupled enzymatic reaction.

C) aromatic amino acids.
D) an antibody that binds the protein of interest.
E) a catalytic antibody.
Ans: D
Level of Difficulty: Easy
Section: 5.1.A
Learning objective: Polypeptide Diversity
11. Which physical characteristic is not commonly used in protein separation?
A) solubility
B) stereochemistry
C) size
D) charge
E) polarity
Ans: B
Level of Difficulty: Easy
Section: 6-1. Protein Isolation, 6-2. Solubilities of Proteins, 6-3. Chromatographic
Separations, 6-4. Electrophoresis

12. Adding additional salt to a protein solution can cause:

A) an increase in solubility called salting in
B) a decrease in solubility called salting out
C) protein precipitation from solution
D) all of the above
E) none of the above
Ans: D
Level of Difficulty: Easy
Section: 6-2. Solubilities of Proteins

13. The acronym HPLC stands for:

A) Hydrophobic protein liquid chromatography
B) High performance liquid chromatography
C) Hydrophilic partition liquid chromatography
D) High priced liquid chromatography
E) Hydrostatic process liquid chromatography
Ans: B
Level of Difficulty: Easy
Section: 6-3. Chromatographic Separations

14. A technique that can be used to separate proteins based primarily their surface non-polar
residues is called:
A) Ion-exchange chromatography
B) Gel filtration chromatography
C) Afffinity chromatography
D) Gel electrophoresis
E) Hydrophobic interaction chromatography
Ans: E
Level of Difficulty: Easy
Section: 6-3. Chromatographic Separations

15. SDS-PAGE separates proteins primarily due to differences in:

A) isoelectronic points
B) size
C) polarity

D) solubility
E) sequence
Ans: B
Level of Difficulty: Easy
Section: 6-4. Electrophoresis

16. Which of the following amino acids would be last to elute at high pH from an anionexchange column?
A) lysine
B) alanine
C) glutamic acid
D) asparagine
E) glycine
Ans: C
Level of Difficulty: Moderate
Section: 6-3. Chromatographic Separations