There are a number of reducing agents that are available at Gold Bio: DTT (dithiothreitol), DTE (dithioerythritol), L-glutathione (GSH) and TCEP (Tris (2-Carboxyethyl) phosphine hydrochloride). By definition, reducing agents are elements or compounds that donate an electron to an oxidizer compound, hence a compound may be “reduced” (lose an electron) to create an “oxidized” state and the reaction can typically be reversed by “oxidizing” (donating an electron) a compound back into its “reduced” state. At the protein level, reducing agents are often critical in cleaving the disulfide bonds between cysteine amino acids. This effect is probably most commonly experienced by the non-scientist in the form of a chemical hair perm, in which hair is chemically straightened (reduction of the disulfide bonds that make hair curly), and later curled (oxidized to reinstate those disulfide bonds) into a specific structure. Because of the difficulty of reducing disulfide bonds that are deeply buried, reduction is often done in the presence of high temperatures or denaturants such as guanidine-HCl or urea to help linearize the protein so that the reducing agent can work more effectively.

Dithiothreitol (DTT) is a particularly strong reducer because once in its oxidized state, it forms a very stable ring structure with an internal disulfide bond which makes it harder to oxidize back to its reduced state again. DTT can also be used for reducing the disulfide bridge of the cross-linker N,N′-bis(acryloyl) cystamine to break apart the matrix of a polyacrylamide gel. DTT is a nearly 7-fold stronger reduction agent than βME (β-mercaptoethanol) and does not have the disadvantages of the repulsive smell or toxicity of βME.

But everything is multifunctional. Among many other applications, DTT is commonly used as a cell-free measure of the oxidative potential of particles in that redox-active chemicals in particulate matter (PM) oxidize the added DTT to its disulfide form and the linear rate of DTT loss is used as a measure of the oxidative capacity of the PM. And due to its protein denaturing ability, it has been widely used in microbiology labs for liquefying specimens from the respiratory tract and has been reported to reduce staphylococcyl biofilm. It has been shown to putatively remove that bacterial biofilm from the surface of prosthetic materials, making it a useful tool in the screening and treatment of PJI (Prosthetic Joint Infection). And perhaps of greater importance (to some), it has been used to help measure the “limit dextrinase” (a debranching enzyme found in some key cereals) activity during barley malting in scotch whiskey production.

As a reducing agent, DTT has proven to be useful both in simple protein studies as well as an ever widening range of other applications outside of the typical ‘proteomic box’ in industry and medicine.

Charrier, J. G., and C. Anastasio. "On dithiothreitol (DTT) as a measure of oxidative potential for ambient particles: evidence for the importance of soluble transition metals." Atmospheric Chemistry and Physics Discussions 12 (2012): 11317-11350.

Drago, Lorenzo, et al. "Does Dithiothreitol Improve Bacterial Detection from Infected Prostheses? A Pilot Study." Clinical Orthopaedics and Related Research® (2012): 1-11.

Wu, Xiaoqian, Yu Wang, and Liang Tao. "Sulfhydryl compounds reduce Staphylococcus aureus biofilm formation by inhibiting PIA biosynthesis." FEMS Microbiology Letters 316.1 (2011): 44-50.

Walker, J. W., et al. "The survival of limit dextrinase during fermentation in the production of Scotch whisky." Journal of the Institute of Brewing 107.2 (2012): 99-106.

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