Lectin Purification by Ion Exchange Chromatography (CAT#: STEM-MB-1299-LGZ)

The principle of separation is thus by reversible exchange of ions between the target ions present in the sample solution to the ions present on ion exchangers. In this process, two types of exchangers i.e., cationic and anionic exchangers can be used.
Cationic exchangers possess negatively charged group, and these will attract positively charged cations. These exchangers are also called “Acidic ion exchange” materials, because their negative charges result from the ionization of acidic group.
Anionic exchangers have positively charged groups that will attract negatively charged anions. These are also called “Basic ion exchange” materials.
Ion exchange chromatography is most often performed in the form of column chromatography. However, there are also thin-layer chromatographic methods that work basically based on the principle of ion exchange.

For the purification of lectins.

1. Equilibration
The first step is the equilibration of the stationary phase. When equilibrium is reached, all stationary phase charged groups are bound with exchangeable counterions, such as chloride or sodium. The pH and ionic strength of the start buffer are selected to ensure proteins of interest bind to the medium.
2. Sample Application and Wash
The goal in the second step is to bind the target molecule(s) and wash out all unbound material.
3. Elution
The proteins with the lowest net charge at the selected pH will be the first ones eluted from the column as ionic strength increases. Similarly, the proteins with the highest charge at a certain pH will be most strongly retained and will be eluted last.
4. Regeneration
A final wash with high ionic strength buffer regenerates the column and removes any molecules still bound. The column is then re-equilibrated in start buffer before starting the next run.

• Sample containing lectin