Molecular cloning is a commonly used technique for the generation of recombinant proteins. The gene for a protein of interest is cloned into a vector, or plasmid, in frame with the gene for a protein tag. These tags are used to purify expressed protein by established methodologies. The most common tags in use consist of the six histidine motif (6XHis), purified on a nickel metal chelating column; the glutathione S-transferase (GST) tag, purified with a glutathione resin; and the calmodulin binding peptide (CBP) tag, isolated with calmodulin resin.
The Recombinant Protein Purification kit teaches students the basic principles of purifying recombinant proteins and has a hands-on lab activity that teaches students the purification of a tagged protein. The recombinant protein supplied is a light emitting protein that allows for easy detection of the purified protein, using UV light. Students will learn about protein tags and affinity chromatography.
This lab activity teaches the following:
- Strategies used in the purification of recombinant proteins.
- Purification of a recombinant protein.
- Purification of recombinant tagged protein.
Please refer to the Product Protocol for additional equipment needed.