1. Biochemical Assay Reagents
  2. DTNB

DTNB (Synonyms: Ellman’s Reagent)

Cat. No.: HY-15915 Purity: 98.35%
Handling Instructions

DTNB (Ellman’s Reagent) is a chemical used to quantify the number or concentration of thiol groups.

For research use only. We do not sell to patients.

DTNB Chemical Structure

DTNB Chemical Structure

CAS No. : 69-78-3

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10 mM * 1 mL in DMSO USD 55 In-stock
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Based on 1 publication(s) in Google Scholar

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Description

DTNB (Ellman’s Reagent) is a chemical used to quantify the number or concentration of thiol groups[1].

In Vitro

DTNB reacts with the free sulfhydryl side chain of cysteine to form an S-S bond between the protein and a thionitrobenzoic acid (TNB) residue[1].
The main advantage of DTNB over alternative reagents (e.g., N-ethylmaleimide or iodoacetamide) is in the selectivity of this reagent and in the ability to follow the course of the reaction spectrophotometrically[1].
Modification of Enzyme with DTNB:
Modification of the SH groups of the enzyme is carried out by reacting 10 μL of 10 mM DTNB solution (about a 20-fold molar excess) at room temperature with 0.6 mL of enzyme solution (0.807 mg/mL) in 80 mM phosphate buffer, pH 8.0, containing 1 mM EDTA, which has been dialyzed previously against the same buffer solution for 24 h. The number of SH groups is estimated from the increase of absorbance at 410 nm using a molar extinction coefficient of 13,600 M-1cm-1 for thionitrobenzoate ions liberated.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

396.35

Formula

C₁₄H₈N₂O₈S₂

CAS No.

69-78-3

SMILES

O=C(O)C1=C([N+]([O-])=O)C=CC(SSC2=CC(C(O)=O)=C([N+]([O-])=O)C=C2)=C1

Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 6 months
-20°C 1 month
Solvent & Solubility
In Vitro: 

DMSO : ≥ 150 mg/mL (378.45 mM)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.5230 mL 12.6151 mL 25.2302 mL
5 mM 0.5046 mL 2.5230 mL 5.0460 mL
10 mM 0.2523 mL 1.2615 mL 2.5230 mL
*Please refer to the solubility information to select the appropriate solvent.
In Vivo:
  • 1.

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% saline

    Solubility: ≥ 2.08 mg/mL (5.25 mM); Clear solution

  • 2.

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in saline)

    Solubility: ≥ 2.08 mg/mL (5.25 mM); Clear solution

  • 3.

    Add each solvent one by one:  10% DMSO    90% corn oil

    Solubility: ≥ 2.08 mg/mL (5.25 mM); Clear solution

*All of the co-solvents are provided by MCE.
References

Purity: 98.35%

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The molarity calculator equation

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2

Keywords:

DTNBEllman’s ReagentOthersInhibitorinhibitorinhibit

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