GST Agarose

GST Agarose

Glutathione Agarose is an effective way for purification of recombinant Glutathione-S-transferase fusion proteins from cellular lysates. The product is prepared by coupling Glutathione through it's central sulphydryl group by a linker, which minimizez steric hindrance, to activated agarose.

The Glutathione-S-transferase (GST) are a family of enzymes that catalyze the addition of the tripeptide glutathione to endogenous and xenobiotic substrates which have electrophilic functional groups:

They play an important role in the detoxification and metabolism of many xenobiotic and endobiotic compunds, because their addusts have increased solubility in water and are subsequently enzymatically degrade to mercapturates and excreted. Glutathione-S-transferase are dimeric with subunits of 25kDa molecular weight.

In general the GST gene fusion system is an intergrated system for expression, purification and detection of fusion proteins expressed in baterial, yeast, mammalian and insect cells. The sequence encoding the GST protein is incorporated into an expression vector, generally upstream of the multi-cloning site. The sequence encoding the protein of interest is then cloned into this vector. Induction of the vector results in expression of the fusion protein- the protein of interest fused to the GST protein. The fusion protein can then be released from the cells and purified.

Purification of the fusion protein is facilitated by the affinity of the GST protein and glutathione residues, which are coupled to Biontex GST Agarose resin. The expression fusion protein product is brought in contact with the resin and will bind to the glutathione-agarose complex. Now, all other non-specific proteins can be washed off. the fusion protein can then be released from the resin using a mild elution buffer including glutathione or by cleveage from fusion protein by using a specific protease, which cleave specific sites between GST and the protein of interest.

Fusion Proteins can also be detected with a number of GST antibodies.

Last modified: 2008-05-14 16:46:46