TMB
Based on 8 publication(s) in Google Scholar
TMB is a non-toxic, non-mutagenic, colorless dye that is also used as a chromogenic agent in enzyme detection systems based on horseradish peroxidase (HRP). TMB can function as a photoacoustic probe for reactive oxygen and nitrogen species (ROS/RNS), reacting with hypochlorite, hydrogen peroxide, singlet oxygen, and nitrogen dioxide to produce blue oxidation products. TMB can detect the singlet oxygen produced by the outer membrane of Escherichia coli ATCC 25922.
For research use only. We do not sell to patients.
- Purity : 99.98%
- CAS No.: 54827-17-7
- Formula: C16H20N2
- Molecular Weight:240.34
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Storage:
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Help & FAQs
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PBS Buffer (1×)
Publications Citing Use of MedChemExpress (MCE) TMB
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Customer Validation & Images
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Bio/Physico-chemical Assay
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Bio/Physico-chemical Assay
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Bio/Physico-chemical Assay
Biological Activity
Description
In Vitro
TMB can tightly bind to Escherichia coli ATCC 25922 cells instead of being internalized to detect singlet oxygen produced by the outer membrane[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
Chemical Information
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CAS No. 54827-17-7
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Appearance Solid
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Molecular Weight 240.34
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Formula C16H20N2
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Color Off-white to light yellow
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SMILES
NC1=C(C)C=C(C2=CC(C)=C(N)C(C)=C2)C=C1C
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Synonyms
BM blue; Sure Blue TMB
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Publications (8)
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Journal Impact Factor
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Most Recent
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TMB purchased from MedChemExpress. Usage Cited in: Chem Eng J. 2025 Nov 24;526:171230.
A diagram showing the chemical reaction formula of TMB assay, MB assay, and DTNB assay.
TMB purchased from MedChemExpress. Usage Cited in: Chem Eng J. 2025 Nov 24;526:171230.
The results of TMB assay.
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J Control Release
Synergistic photothermal-sonodynamic therapy for antibacterial and immune reprogramming in chronic osteomyelitis. [Abstract]2025 May 10:381:113612. PMID: 40073945 -
TMB purchased from MedChemExpress. Usage Cited in: Nano Res. 2026 Feb 6.
Ov-ZBC showed strong characteristic peaks, indicating its higher photocatalytic ROS generation capability. The results of TMB and MB chromogenic reactions shown in Fig.
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Int J Biol Macromol
Construction and characterization of a novel single-chain variable fragment against Bacillus thuringiensis vegetative insecticidal protein. [Abstract]2025 Sep 14;328(Pt 1):147690. PMID: 40957516 -
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J Invertebr Pathol
Characterization of the individual domains of the Bacillus thuringiensis Cry2Aa implicates Domain I as a possible binding site to Helicoverpa armigera. [Abstract]2024 Jul:205:108129. PMID: 38754546 -
Solvent & Solubility
In Vitro:
DMSO : 25 mg/mL (104.02 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Protocols
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ROS/oxidative-stress fluorescent staining
ROS/oxidative-stress fluorescent staining uses cell-permeant fluorogenic probes that become fluorescent after oxidation inside cells or tissues; commonly used examples include DCFH-DA/DCFDA for broad cellular oxidant detection, DHE for superoxide-related signal detection, MitoSOX for mitochondrial superoxide-related signal detection, and CellROX probes for oxidative-stress-associated fluorescence readouts. The assay detects probe oxidation rather than a single ROS species unless the probe and analysis method have been chemically validated for that species. DCFH-DA enters cells, is deacetylated by intracellular esterases to DCFH, and produces fluorescent DCF after oxidation, so the readout is used as an operational measure of total cellular oxidative stress rather than a species-specific ROS measurement. DHE and MitoSOX can report superoxide-related oxidation, but red fluorescence alone can include non-specific ethidium-like oxidation products; HPLC or optimized spectral approaches are
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Genotoxicity/Mutagenicity Study
The bacterial reverse mutation assay detects point mutations that restore amino-acid prototrophy in auxotrophic Salmonella typhimurium or Escherichia coli tester strains; after exposure to a test article, mutagenic activity is read out as an increased number of revertant colonies on minimal agar compared with the vehicle control. The assay uses tester strains with different mutation targets so that base-substitution and frameshift mutagens can be detected, and testing is performed with and without exogenous mammalian metabolic activation because some chemicals require biotransformation to become mutagenic.
Purity & Documentation
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Data Sheet (272 KB)
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SDS (557 KB)
- English - EN (557 KB)
- Français - FR (557 KB)
- Deutsch - DE (557 KB)
- Norwegian - NO (557 KB)
- Español - ES (557 KB)
- Swedish - SV (557 KB)
- Italian - IT (557 KB)
- Korean - KR (557 KB)
- Portuguese - PT (557 KB)
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Handling Instructions (2659 KB)
References
[1]. Bresolí-Obach R, et al. Tetramethylbenzidine: An Acoustogenic Photoacoustic Probe for Reactive Oxygen Species Detection. Sensors (Basel). 2020 Oct 21;20(20):5952. [Content Brief]
[2]. Frey A, et al. A stable and highly sensitive 3,3',5,5'-tetramethylbenzidine-based substrate reagent for enzyme-linked immunosorbent assays. J Immunol Methods. 2000 Jan 13;233(1-2):47-56. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 4.1608 mL | 20.8039 mL | 41.6077 mL | 104.0193 mL |
| 5 mM | 0.8322 mL | 4.1608 mL | 8.3215 mL | 20.8039 mL | |
| 10 mM | 0.4161 mL | 2.0804 mL | 4.1608 mL | 10.4019 mL | |
| 15 mM | 0.2774 mL | 1.3869 mL | 2.7738 mL | 6.9346 mL | |
| 20 mM | 0.2080 mL | 1.0402 mL | 2.0804 mL | 5.2010 mL | |
| 25 mM | 0.1664 mL | 0.8322 mL | 1.6643 mL | 4.1608 mL | |
| 30 mM | 0.1387 mL | 0.6935 mL | 1.3869 mL | 3.4673 mL | |
| 40 mM | 0.1040 mL | 0.5201 mL | 1.0402 mL | 2.6005 mL | |
| 50 mM | 0.0832 mL | 0.4161 mL | 0.8322 mL | 2.0804 mL | |
| 60 mM | 0.0693 mL | 0.3467 mL | 0.6935 mL | 1.7337 mL | |
| 80 mM | 0.0520 mL | 0.2600 mL | 0.5201 mL | 1.3002 mL | |
| 100 mM | 0.0416 mL | 0.2080 mL | 0.4161 mL | 1.0402 mL |
TMB Related Classifications
HY-15930 Related Classifications
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