IDA-His-tagged Protein Purification Kit

Immobilized metal ion affinity chromatography (IMAC) is a rapid affinity purification chromatography where the his-tagged protein are separated based on their affinity for Ni²⁺ or Co²⁺that have been immobilized by a chelator to a solid matrix such as beaded agarose or column. At pH 7-8, his tagged protein will bind to Ni²⁺ or Co²⁺. The binding reaction with the tagged protein is affected by various independent variable factors such as pH, temperature, salt type, salt concentration, immobilized metal and ligand density, and protein size. The bound protein is eluted by a decreasing pH gradient, increasing imidazole concentration, or adding an EDTA chelator in a buffer. This technique is an ideal tool for capturing and purification of his-tagged recombinant protein in a quick, inexpensive, and straightforward manner.