Ellman’s Reagent, 10 mM DTNB Solution (5,5′-Dithiobis(2-nitrobenzoic acid)) BZ355
Product Code: BZ355
Manufacturer: Biochemazone
Available Pack Sizes: 100 mL, 200 mL, 500 mL, 1 L
Custom Bulk Quantities Available
Product Overview
Biochemazone Ellman’s Reagent BZ355 is a ready-to-use 10 mM solution of DTNB (5,5′-dithiobis(2-nitrobenzoic acid)) for measuring free thiol (–SH) groups in proteins, peptides and small molecules such as cysteine and glutathione.
DTNB reacts with a free thiol to release one molecule of the yellow TNB2− anion, which is read at 412 nm (molar extinction coefficient 14.15 mM−1 cm−1). The 10 mM stock is simply diluted into your assay buffer – no weighing of DTNB powder. The same chemistry underpins the Ellman acetylcholinesterase assay, where acetylthiocholine hydrolysis is followed with DTNB.
Each batch is prepared under Biochemazone’s ISO 9001 / GMP quality system.
Key Features & Benefits
✅ 10 mM DTNB Stock – Ready to dilute, no weighing
✅ Fast Read at 412 nm – 2-minute reaction, yellow TNB product
✅ Quantitative – ε412 = 14.15 mM−1 cm−1
✅ Plate or Cuvette – Works in microplates and spectrophotometers
✅ AChE Assays – Classic Ellman cholinesterase method
✅ 100 mL to 1 L – Plus custom bulk volumes
✅ Quality-Controlled – Consistent batch-to-batch performance
✅ Made in Canada – Manufactured under ISO 9001 / GMP quality systems
Applications
🧪 Protein & Peptide Chemistry
• Free-thiol (cysteine) quantification
• Checking disulfide reduction
• Monitoring thiol–maleimide and PEGylation reactions
🧬 Biochemistry & Pharmacology
• Acetylcholinesterase and butyrylcholinesterase activity (Ellman method)
• Glutathione and cysteine measurement
• Enzyme inhibitor screening
🔬 Analytical Chemistry
• Thiol content of foods, drugs and materials
• Teaching laboratories
Usage Areas
- Protein chemistry and bioconjugation laboratories
- Pharmacology and toxicology laboratories
- Neuroscience (cholinesterase) research groups
- Food and pharmaceutical analysis laboratories
- Academic and teaching laboratories
Composition (per litre)
| Component | Concentration | Amount per litre |
|---|
| DTNB (5,5′-dithiobis(2-nitrobenzoic acid), CAS 69-78-3) | 10 mM | 3.96 g |
| Solvent | — | to 1 L (as stated on the CoA) |
Molecular weight of DTNB: 396.3 g/mol. Dilute 100-fold in 0.1 M Tris-HCl, pH 7.5 for a 0.1 mM working solution.
Assay Protocol (1 mL cuvette)
| Step | What to do |
|---|
| 1. Working solution | Dilute BZ355 100-fold with 0.1 M Tris-HCl, pH 7.5, to give 0.1 mM DTNB. |
| 2. Sample | Mix 950 µL working solution with 50 µL sample (thiol below 0.5 mM). |
| 3. Blank | Mix 950 µL working solution with 50 µL 0.1 M Tris-HCl, pH 7.5. |
| 4. Incubate | 2 minutes at room temperature. |
| 5. Read | Measure absorbance at 412 nm against the blank. |
| 6. Calculate | mM –SH = A412 ÷ (path length × 14.15) × 20 × sample dilution factor. |
The factor 20 accounts for 50 µL sample in a 1 mL assay. Adapt volumes for microplates or use a cysteine standard curve.
Typical Properties
- Appearance: Clear, pale yellow liquid
- DTNB concentration: 10 mM
- Detection: Absorbance at 412 nm
- Light-sensitive: keep protected from light
Available Formulations
- Ellman’s reagent 10 mM DTNB (this product, BZ355)
- 1 M Tris-HCl pH 7.4 (BZ217)
- 1 M Tris-HCl pH 8.0, sterile (BZ214)
- 0.1 M phosphate buffer pH 7.4 (BZ204)
Storage, Shelf Life & Transportation
- Storage Temperature: 2–30 °C, protected from light
- Shelf Life: 1 year for unopened bottles
- Transportation: Room temperature (ambient shipping)
- Version with Preservatives: Available on request
- Powder Version: Available on request
- Keep tightly closed; prepare working dilutions fresh
- A strong yellow colour in the stock indicates degradation – check against a blank
Customization Options
- Other DTNB concentrations
- Ready-to-use 0.1 mM working solution in Tris-HCl pH 7.5
- DTNB powder and cysteine standards
- Bulk containers
Ordering Information
BZ355 is available in standard laboratory sizes or OEM/bulk supply formats.
📄 MSDS, Technical Datasheet, and Certificate of Analysis (CoA) available upon request
Email: info@biochemazone.com
Website: www.biochemazone.com
⚠️ For laboratory research use only. Not for human or animal injection or therapeutic use.