Dithiothreitol, or DTT for short. It is a small molecule organic reducing agent with the chemical formula C4H10O2S2. In the reduced state, it is a linear molecule, and after oxidation, it becomes a six-membered ring structure containing disulfide bonds. The name dithiothreitol is derived from threose (a four-carbon monosaccharide). The isomer of DTT is dithioerythritol (DTE), which is the C3-heterotropic isomer of DTT.
Chemical formula C4H10O2S2
Molar mass 154.253 g·mol−1
Appearance White solid
Melting point 42-43 °C
Boiling point 125-130 °C (at 2 mmHg pressure)
Solubility in water is soluble
One of the uses of DTT is as a reducing agent and a deprotective agent for sulfhydrylated DNA. Sulfhydrated DNA terminal sulfur atoms tend to form dimers in solution, especially in the presence of oxygen. This dimerization greatly reduces the efficiency of some coupling reaction experiments (such as DNA fixation in biosensors); By adding DTT to DNA solution and removing it after a period of reaction, the dimerization of DNA can be reduced.
DTT is also commonly used to reduce disulfide bonds in proteins, and can be used to prevent the formation of protein intramolecular or intermolecular disulfide bonds between the cysteines in proteins. However, DTT is often unable to reduce the disulfide bonds embedded in the protein structure (not accessible to solvents), and the reduction of such disulfide bonds often requires denaturing the protein (high temperature heating or the addition of denaturing agents, such as 6M guanidine hydrochloride, 8M urea or 1% SDS). On the contrary, the depth of the embedding degree can be judged according to the different reduction rate of disulfide bond in the presence of DTT.
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