Overview
Dithiothreitol (DTT) is a small-molecule reducing agent widely used in biochemistry and molecular biology. It was first synthesized by Walter Cleland in 1964 as a safer alternative to β-mercaptoethanol. DTT’s primary function is to cleave disulfide bonds in proteins, making it indispensable for protein structure analysis and manipulation. Its stability in aqueous solutions and low toxicity compared to other reductants have made DTT a standard reagent in laboratories. It is particularly valued for its role in maintaining protein solubility during electrophoresis and in protecting enzymes from oxidative inactivation.
Physical and Chemical Properties
DTT is a white, crystalline solid with a molecular weight of 154.25 g/mol. It melts at 42-44°C and is highly soluble in water, ethanol, and acetone. The compound’s reducing power stems from its two thiol groups, which readily donate hydrogen atoms to break disulfide bridges. A key limitation is its oxidation sensitivity; DTT decomposes in air, forming an inactive cyclic disulfide. To mitigate this, solutions are often prepared fresh or stored under inert gas. Its pKa of ~9.2 ensures effective reduction across a broad pH range, though activity declines in highly acidic conditions.
Main Applications
In protein research, DTT is used to denature proteins by reducing disulfide bonds, enabling techniques like SDS-PAGE. It also facilitates protein refolding by preventing incorrect bond formation during purification. Pharmaceutical applications include its use in antibody production and as a stabilizing agent for sensitive enzymes. Beyond life sciences, DTT serves as a reducing agent in chemical synthesis, particularly for creating metal nanoparticles. Its ability to maintain sulfhydryl groups in a reduced state is critical for industrial biocatalysis and diagnostic assay development.
Safety and Storage
DTT is classified as an irritant and requires handling with gloves and eye protection. Inhalation of dust should be avoided, and work should be conducted in fume hoods if large quantities are used. Spills can be neutralized with dilute hydrogen peroxide. For storage, DTT is best kept in airtight containers under nitrogen or argon at 2-8°C. Lyophilized powder is more stable than solutions, which typically degrade within days at room temperature. Industrial users should prioritize suppliers offering stabilized formulations for extended shelf life.
B2B Procurement Guide
When sourcing DTT, verify the CAS number (3483-12-3) to ensure product authenticity. Research-grade purchases (≥99% purity) are common for laboratories, while industrial users may opt for technical-grade batches. Bulk pricing is typically available for orders above 1 kg. Key procurement considerations include packaging (amber glass vials for light-sensitive stability), certificates of analysis, and supplier compliance with ISO or GMP standards. Lead times can vary; specialty grades may require 2-4 weeks for custom preparation.
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