Preparation of 1 M Dithiothreitol (DTT) solution

Overview

  • DTT is used to stabilize enzymes and other proteins which possess free sulfhydryl groups.
  • Formula : C4H10O2S2
  • Molecular weight : 154.25

Requirement

  • Reagents
    • DL-Dithiothreitol (C4H10O2S2) (Molecular weight = 154.25)
    • Deionized / Milli-Q water
  • Equipments and disposables
    • Measuring cylinder
    • Conical flask / Beaker
    • Magnetic stirrer

Composition

  • 1 M Dithiothreitol

Objective

  • Preparation of 25 ml of 1 M Dithiothreitol solution

Preparation

Step 1: To prepare 25 ml aqueous solution of 1M DTT, weigh out 3.86 gram of DTT (Molecular weight = 154.25). Transfer it to 50 ml beaker/conical flask. Add 20 ml deionized / Milli-Q water. Mix until all DTT is dissolved completely.

Tips:

  • One can use manual shaking using a glass pipette to mix the ingredients. Magnetic stirrer makes the dissolving process easy and convenient.

Precautions:

  • Do not dissolve in 25 ml of deionized / Milli-Q water. In most cases, solution volume increases when the large amount of solute dissolves in solvent. see solution preparation

Step 2: Adjust the volume to 25 ml with deionized / Milli-Q water. Mix it again.

Step 3 (Optional): One can sterilize the solution by passing through 0.22 μ filter unit under aseptic condition (inside laminar flow hood).

Precautions:

  • Do not autoclave the solution. Autoclaving will destroy the DTT.

Storage

Solution should be stored at -20°C.

Tips:

  • It is recommended to store the solutions as small aliquots.

Precautions:

  • Avoid repeated handling and multiple freeze/thaw cycles.

Applications

  • Enzyme stabilization
  • Enzymatic reactions
  • Protein electrophoresis
To prepare Rubidium chloride solution of different concentration and volume, follow the table.
Conc. / Volume 5 ml 10 ml 25 ml 50 ml 100 ml 200 ml
10 mM 0.0077 g 0.015 g 0.0386 g 0.077 g 0.154 g 0.309 g
100 mM 0.077 g 0.154 g 0.386 g 0.77 g 1.54 g 3.085 g
0.5 M 0.385 g 0.77 g 1.93 g 3.86 g 7.71 g 15.43 g
1 M 0.77 g 1.54 g 3.86 g 7.71 g 15.43 g 30.85 g