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Application, precautions and preservation methods of dithiothreitol (DTT)

2019-04-10 来源:亚科官网
Dithiothreitol (DTT) is a small molecule organic reducing agent with the chemical formula C4H10O2S2. In the reduced state, it is a linear molecule, and after being oxidized, it becomes a six-membered ring structure containing a disulfide bond. DTT discovered in the middle of the last century that this small molecule and water-soluble reagent can completely reduce disulfide, protect sulfhydryl groups from oxidation, and preserve monoterpenes in a reduced state. As an effective reducing agent, it is mainly used in clinical and experimental fields, and can also be used in the treatment of certain diseases, such as cystine disease or medical conditions caused by ionic or metal toxicity.
Application of dithiothreitol
One of the uses of DTT is as a reducing agent and deprotecting agent for thiolated DNA. The thiolated DNA terminal sulfur atom tends to form a dimer in solution, especially in the presence of oxygen. This dimerization greatly reduces the efficiency of some coupling reaction experiments (such as the immobilization of DNA in biosensors); while adding DTT to the DNA solution and removing it after a period of reaction, the dimerization of DNA can be reduced.
DTT is also often used for the reduction of disulfide bonds in proteins and can be used to prevent intramolecular or intermolecular disulfide bonds of proteins formed between cysteines in proteins. However, DTT often cannot reduce the disulfide bonds embedded in the protein structure (solvent unreachable). The reduction of such disulfide bonds often requires denaturation of the protein (heating at high temperature or adding a denaturing agent such as 6M guanidine hydrochloride, 8M urea or 1% SDS). On the contrary, according to the difference in the rate of reduction of disulfide bonds in the presence of DTT, the degree of embedding can be judged.
Precautions of Dithiothreitol
DTT is less stable due to its ease of oxidation by air; however, cryopreservation or treatment in an inert gas can extend its useful life. Since the nucleophilicity of protonated sulfur is low, the effective reduction of DTT decreases as the pH decreases, so the pH of the system should be controlled.
Method for preserving dithiothreitol
Store at 2~8 °C in cold storage. Sealed to avoid moisture, heat and direct sunlight.
Related links: Dithiothreitol
Edited by Suzhou Yacoo Science Co., Ltd.