Targeting Tools: Anti-GST

The Glutathione S-Transferase (GST) family of enzymes contains cytosolic, mitochondrial, and microsomal proteins that are capable of multiple reactions with a multitude of substrates, both endogenous and xenobiotic. GST catalyzes the conjugation of reduced glutathione which is useful in the detoxification of endogenous compounds such as peroxidized lipids, as well as the metabolism of xenobiotics. GST is used to create the GST gene fusion system in which the GST moiety is used to detect and purify proteins of interest. This GST-fusion protein can then be purified from cell extracts via its high affinity for glutathione. Fusion proteins offer an important biological assay for direct protein-to-protein interactions.

Advanced Targeting Systems announces the release of a new rabbit polyclonal antibody to GST (unlabeled, Cat. #AB-45ap). It is also available labeled with HRP (Cat. # AB-45AP-HRP). Both are useful for the detection of GST fusion proteins.

Cultures of E. coli transformed with a vector expressing a GST-Cre recombinase fusion protein were grown under varying conditions. Soluble protein was extracted from two different clones and expression compared between soluble and insoluble fractions on SDS-PAGE which was then transferred to a PVDF membrane. Membrane was incubated with anti-GST-HRP overnight at a 1:500 dilution.

Lane 1: Page Ruler 5 µl
Lane 2: 200 ng GST-Cre
Lane 3: Supernate 20 µg
Lane 4: Supernate 20 µg
Lane 5: Pellet 5 µl
Lane 6: Pellet 5 µl