Preparation DTT Stock Solution
This guide describes the preparation of DTT Stock Solution at a defined molarity for laboratory use.
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Stock Solution Preparation Guide – DTT (Dithiothreitol)
Dithiothreitol (DTT), also known as Cleland’s reagent, is a commonly used reducing agent in protein biochemistry. It reduces disulfide bonds (–S–S–), stabilizes enzymes, and protects functional thiol groups in proteins. Aqueous stock solutions are best prepared fresh due to susceptibility to oxidation.
Chemical Information
| Name | Molecular Formula | Molecular Weight | CAS |
|---|---|---|---|
| DTT (DL-Dithiothreitol) | C₄H₁₀O₂S₂ | 154.25 g/mol | 3483-12-3 |
Applications
- Reduction of protein disulfide bonds
- Enzyme stabilization
- SDS-PAGE and Western blot sample preparation
Solubility and Handling
- DTT is a white crystalline solid that is highly soluble in water, forming clear solutions.
- Prepare stock solutions in nuclease-free water; air oxidation reduces activity over time.
- Avoid autoclaving DTT solutions as heat degrades the reducing agent.
Stock Solution Preparation
To prepare a 1 M DTT stock solution, calculate the required mass from the molecular weight:
Required mass (g) = Molecular Weight × Volume (L) × Concentration (mol/L)
For 10 mL 1 M solution: 154.25 × 0.010 × 1 = 1.54 g
For 10 mL 1 M solution: 154.25 × 0.010 × 1 = 1.54 g
- Weigh 1.54 g of DTT powder.
- Transfer to a clean tube and add ~8–9 mL of nuclease-free water.
- Mix until completely dissolved.
- Adjust the volume to 10 mL with nuclease-free water.
- Optionally sterilize with a 0.22 µm filter if required.
- Aliquot into single-use volumes and store at −20 °C.
Storage and Stability
- Store solid DTT desiccated at 2–8 °C for long-term stability.
- Stock solutions are best stored at −20 °C in aliquots to minimize oxidation.
- Aqueous DTT is prone to oxidation; prepare fresh working solutions for critical experiments.
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