Hydrophobic chromatography is based on the fact that protein molecules can have extensive hydrophobic regions. These hydrophobic regions, in media favoring hydrophobic interactions, such as an aqueous solution with high salt concentration, can bind to hydrophobic ligands coupled to an uncharged column matrix. Elution is brought about by decreasing the salt concentration and in some cases decreasing the solvent polarity with PEG, non-ionic detergents, denaturants, urea or chaotropic ions.
The Hydrophobic Chromatography kit is designed to teach students the basic principle of hydrophobic chromatography utilizing a hydrophobic enzyme. The use of the enzyme allows purification followed by a simple enzyme assay to detect the fractions that contain the enzyme. This lab activity involves preparation of a crude protein extract and running hydrophobic chromatography to isolate the enzyme.
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 hydrophobic chromatography lab activity.
- The principle of hydrophobic chromatography.
- Immobilization of protein on hydrophobic columns.
- Factors influencing binding and elution of proteins on hydrophobic columns.
- Low speed centrifuge for 1.5 - 2 ml tube
- Waterbath or beaker & thermometer