Biotechniques | Basics of Making His-Tags & Nickel Affinity Chromatography

The video tutorial explains nickel affinity chromatography, a technique used to separate proteins, particularly those engineered with a his tag. It covers the process of adding his tags to proteins using plasmids and E. Coli, and details the steps of nickel affinity chromatography, including binding, washing, and elution using imidazole. The tutorial highlights the importance of this method in molecular biology and biochemistry for protein purification.

What is the primary purpose of nickel affinity chromatography in biochemistry?

• To synthesize amino acids

• To isolate specific proteins

• To separate DNA strands

• To measure enzyme activity

What is a his tag composed of?

• Six glycine residues

• Six lysine residues

• Six histidine residues

• Six alanine residues

Which organism is commonly used to express proteins with a his tag?

• Bacillus subtilis

• Escherichia coli

• Saccharomyces cerevisiae

• Pseudomonas aeruginosa

What is the role of nickel NTA agarose beads in the chromatography process?

• To degrade unwanted proteins

• To bind proteins with a his tag

• To stabilize the protein structure

• To provide a fluorescent signal

What is used to elute the protein of interest from the nickel NTA beads?

• Acetic acid

• Ethanol

• Imidazole

• Sodium chloride

Why is imidazole effective in displacing the his tag from nickel?

• It binds to DNA

• It is a strong acid

• It is a reducing agent

• It has a similar structure to histidine

What is the main advantage of using nickel affinity chromatography?

• It works with all types of proteins

• It requires no special equipment

• It can purify proteins with high specificity

• It is the fastest method available