| Description | GST family of enzymes comprises a long list of cytosolic, mitochondrial, and microsomal proteins that are 45-55 kDa (dimer form) size and are capable of multiple reactions with a multitude of substrates, both endogenous and xenobiotic. GST catalyses the conjugation of reduced glutathione meaning the sulfhydryl group, to electrophilic centers on a wide variety of substrates. This activity is useful in the detoxification of endogenous compounds such as peroxidised lipids, as well as the metabolism of xenobiotics. GST binds toxins and function as transport protein. The soluble Glutathione S-Transferase is a 26 kDa protein which occurs as a dimer in all aerobic organisms. Each monomer has two domains, one that binds GSH and is an alpha/beta-structure similar to thioredoxin and the other, all helical, that binds the hydrophobic substrate (1). The GST-fusion protein expression system is a widely used recombinant protein expression system. Fusion proteins also possess GST-enzymatic activity and can undergo dimerization similar to in vivo. The fusion protein can be purified via GST-affinity column chromatography. In most cases, the desired peptides or domains are removed from GST by applying a specific protease that recognizes and cleaves the linker between the protein domain and GST. The technique has been widely used to generate different kinds of proteins for crystallization (3), molecular immunology studies (4), the production of vaccines (5) and studies involving protein-protein (6) and protein-DNA interactions (7). | | Purification Method | GST antibody was purified from rabbit blood serum by protein-A affinity chromatography. | | Formulation | 0.7mg/ml in PBS, pH7.4. | | Storage and stability | For periods up to 1 month store at 4°C, for longer periods of time, store at -20°C. Avoid freeze thaw cycles. | | Application | GST antibody has been tested ELISA and Western blot analysis to assure specificity and reactivity. α-GST reacts with the GST protein in HeLa nuclear extract by Western Blotting. Since application varies, however, each investigation should be titrated by the reagent to obtain optimal results. Recommended dilution range for Western blot analysis is 1:1,000 ~ 10,000. Recommended starting dilution is 1:3,000. For Research use only. | | References: | 1. Ketterer, B. (2001) Chem Biol Interact138(1):27-42
2. Smith, D.B. et al., (1988) Gene 67, 31-34
3. Zan et al., (2001) Gene 281(1-2):1-9
4. Toye, B. et al., (1990) Infect. Immun. 58, 3909-3914
5. Fikrig, E., et al., (1990) Science 250, 553-556
6. Kaelin, W.G. et al., (1991) Cell 64, 521-529
7. Kaelin, W.G. et al., (1991) Cell 65, 1073-1080. |
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