| Size | Price | Stock | Qty |
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| 1g |
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| 5g |
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| Other Sizes |
| Targets |
TMB dihydrochloride targets horseradish peroxidase (HRP) and other peroxidase enzymes, acting as a hydrogen donor in the enzymatic reaction. It participates in the reduction of hydrogen peroxide to water, a process catalyzed by peroxidases, producing a measurable colorimetric change.
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| ln Vitro |
TMB dihydrochloride is a colorless crystalline powder that turns blue in the presence of peroxidase enzymes. It acts as a substrate for HRP, wherein the enzyme catalyzes the oxidation of TMB in the presence of hydrogen peroxide. The substrate produces a soluble end product that is pale blue in color and can be read spectrophotometrically at 370 or 650 nm.
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| ln Vivo |
As a biochemical reagent, TMB dihydrochloride is not typically evaluated for in vivo activity in animal models. Its primary application is in diagnostic and research assays rather than therapeutic in vivo studies.
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| Enzyme Assay |
In vitro enzyme assays using TMB dihydrochloride involve incubating the substrate with HRP-labeled probes or peroxidase enzymes in a buffer system containing hydrogen peroxide. The reaction is stopped with acid (e.g., H₂SO₄) and the absorbance is measured at 450 nm (for the yellow end-product) or 650 nm (for the blue product) to quantify enzyme activity or analyte concentration.
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| Cell Assay |
In vitro cell-based assays using TMB dihydrochloride are typically performed in ELISA or immunohistochemistry formats. Cells or tissue sections are incubated with HRP-conjugated antibodies, followed by addition of TMB substrate solution. Color development is monitored and the reaction is stopped to measure absorbance, allowing quantification of target antigen expression or cellular enzyme activity.
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| Animal Protocol |
In vivo animal studies are not typically performed with TMB dihydrochloride as it is a laboratory reagent rather than a therapeutic agent. It is used in ex vivo analyses of tissue samples from animal studies, where HRP-based detection systems employing TMB visualize target proteins in tissue sections or biological fluids.
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| ADME/Pharmacokinetics |
TMB dihydrochloride has a molecular weight of 313.27 g/mol and formula C₁₆H₂₂Cl₂N₂. It is soluble in water (2 mg/mL) and DMSO (6 mg/mL), but insoluble in ethanol. As a substrate, it is rapidly oxidized by peroxidases and does not exhibit systemic pharmacokinetics as it is not administered in vivo for therapeutic purposes.
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| Toxicity/Toxicokinetics |
TMB dihydrochloride is considered a safe alternative to carcinogenic benzidine. It is classified as a skin and eye irritant; appropriate laboratory safety precautions should be followed. No significant systemic toxicity has been reported as it is not intended for in vivo administration.
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| Additional Infomation |
TMB dihydrochloride is widely used as a chromogenic substrate for HRP in ELISA, immunohistochemistry, and Western blotting applications. It is available in various formulations including TMB-d, BM blue, and Sure Blue TMB. It has been cited in numerous high-impact publications including Nature Medicine for its top-tier quality. Not approved for therapeutic use; intended for research and diagnostic applications only.
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| Molecular Formula |
C16H22CL2N2
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|---|---|
| Molecular Weight |
313.2653
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| Exact Mass |
312.115
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| CAS # |
64285-73-0
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| Related CAS # |
TMB;54827-17-7;TMB monosulfate;54827-18-8
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| PubChem CID |
174033
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| Appearance |
White to off-white solid powder
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| Boiling Point |
368.6ºC at 760 mmHg
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| Melting Point |
≥300 °C
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| Flash Point |
210.8ºC
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| LogP |
6.518
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
20
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| Complexity |
226
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
NYNRGZULARUZCC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H20N2.2ClH/c1-9-5-13(6-10(2)15(9)17)14-7-11(3)16(18)12(4)8-14;;/h5-8H,17-18H2,1-4H3;2*1H
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| Chemical Name |
4-(4-amino-3,5-dimethylphenyl)-2,6-dimethylaniline;dihydrochloride
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ≥ 33 mg/mL (~105.34 mM)
H2O : ~11.11 mg/mL (~35.46 mM) |
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1921 mL | 15.9607 mL | 31.9213 mL | |
| 5 mM | 0.6384 mL | 3.1921 mL | 6.3843 mL | |
| 10 mM | 0.3192 mL | 1.5961 mL | 3.1921 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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