Affinity chromatography is a powerful tool for the purification of specific biomolecules, including proteins. The basic principle is that a biospecific ligand is immobilized to a solid support or resin to which a solution containing the protein of interest is passed over. Ligands are often based on biological functional pairs, such as enzymes and substrate or antigens and antibodies. The specific ligand binds the protein of interest and all non-specific molecules are washed away. The protein is eluted in a specific buffer, either by pH and/or ionic strength shift or by competitively displacement elution.
The Affinity Chromatography kit teaches the basic principle of affinity chromatography utilizing highly specific affinity columns. This lab activity involves preparation of a crude protein extract and running affinity exchange chromatography for isolation of a protein.
Supplied with components needed for hands-on experimentation for six workstations of 4-5 students or 24-30 students. Supplied with Teacher’s Guide and separate Student’s Guides.
- Hands-on affinity chromatography lab activity.
- The principle of affinity chromatography.
- Immobilization of protein on affinity columns.
- Binding and elution of proteins on affinity columns.
- Factors influencing affinity chromatography.