Description

GSTrap HP 1-ml and 5-ml columns are prepacked with Glutathione Sepharose High Performance medium, which provides high-resolution purifications, thus a more concentrated sample. GST-tagged proteins are purified from pretreated bacterial lysates by affinity chromatography using immobilized glutathione, such as Glutathione Sepharose High Performance. GST-tagged proteins are bound to the affinity medium, and impurities are removed by washing with binding buffer GST-tagged proteins are eluted under mild, nondenaturing conditions using reduced glutathione. The purification process preserves protein antigenicity and function. If desired, cleavage of the protein from GST can be achieved using a site-specific protease whose recognition sequence is located immediately upstream from the multiple cloning site on the pGEX plasmids. GST-tagged proteins can be detected using colorimetric or immunological methods.