Dithiothreitol (DTT)
Dithiothreitol (DTT) — A small-molecule reducing agent used to cleave disulfide bonds in peptides and proteins, enabling analysis of individual chains and preventing oxidation.
What Is DTT?
Dithiothreitol (DTT, Cleland's reagent) is a small-molecule reducing agent containing two thiol groups that reduce disulfide bonds in peptides. DTT is widely used in sample preparation for SDS-PAGE, MS, and to maintain Cys residues in the reduced state during assays.
Usage
- Reduction: 1-10 mM DTT at 56°C for 30 min fully reduces peptide disulfides
- Advantage: Complete reduction (DTT is itself oxidized to a cyclic disulfide)
- Limitation: Must be followed by alkylation (IAA) to prevent disulfide re-formation
- Alternative: TCEP is non-thiol reducing agent compatible with maleimide chemistry (DTT is not)
Frequently Asked Questions
What is Dithiothreitol (DTT)?
A small-molecule reducing agent used to cleave disulfide bonds in peptides and proteins, enabling analysis of individual chains and preventing oxidation.
Why is Dithiothreitol (DTT) important in peptide research?
Dithiothreitol (DTT) is a fundamental concept in reagent as it relates to peptide science. It directly influences experimental design, compound characterization, and the reliability of research outcomes across biochemistry and molecular biology disciplines.
Authority Sources
- Dithiothreitol (DTT) on Wikipedia
- Search Dithiothreitol (DTT) on PubChem (NIH)
- Research articles on ScienceDirect