Determine the necessary mass, volume, or concentration for preparing a solution.

Ready-to-use (RTU) for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C,Protected from light Ships Wet ice Check lot-specific COA for exact specifications.
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP, and the addition of a stop solution gives a clear yellow color that absorbs at 450nm.
TMB EASE characteristics are:
1:1 replacement of carcinogenic 2mg OPD
Aqueous based – no organic solvents (REACH Annex XVII compliant)
Low lot-to-lot variation
Product Characteristics
TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP, and the addition of a stop solution gives a clear yellow color that absorbs at 450nm.
TMB EASE is a ready-to-use and stable aqueous formulation. This substrate gives a slow assay reaction and is highly suitable for the direct switch from OPD (ortho-phenylenediamine dichloride) based assays. It does not contain NMP (1-methyl-2-pyrrolidone) making it REACH Restricted Substances List Annex XVII compliant, while ensuring maximal safety during use, and minimal waste problems after use.
Composition & Properties
Ready-to-use substrate: Includes substrate buffer and hydrogen peroxide. No other reagents should be added.
Working Procedure
1. The desired amount of substrate is poured into a sealed container and allowed to reach room temperature in the dark before use.
2. A recommended volume of 100 µl substrate solution per microtiter well is added. Time of development is typically 5-30 minutes. Keep the plate in the dark during incubation.
3. In kinetic assays, the blue color is read directly at 650 nm. For endpoint assays 100 µl (equal volumes) of 0.20 M sulphuric acid is added and the absorbance is read at 450/620 nm. The yellow solution is stable for at least one hour.
Tips & Tricks
• If reduced intensity is required, it is recommended to shorten the time of development or to dilute the conjugate/antibody in the immunoassay further.
• Dilution of the substrate is not recommended.
• In case of incidents of precipitation with very high OD-values, we can recommend the use of the TMB X-tnd (cat. no. T494100).
Handling & Storage
• Store solution at 2-8 °C in the dark.
• Avoid exposure to light, heat and contamination with metal ions or peroxidase.
• Re-dispense only into bottles made of High Density Polyethylene (HDPE), amber color.
• Dispensing guidelines are available upon request.

Comprehensive hazard, handling, storage, and regulatory compliance document.
Download SDS →Lot-specific quality data. Enter your lot number to retrieve the exact COA.
Look up COA →Full quality attributes and acceptance criteria for this grade.
View spec sheet →Find and download the COA for your product by matching the lot number on the packaging.
| Lot Number | Certificate Type | Fecha | Articulo |
|---|---|---|---|
| Certificate of Analysis | Apr 27, 2023 | T494067 | |
| Certificate of Analysis | Apr 27, 2023 | T494067 | |
| Certificate of Analysis | Apr 27, 2023 | T494067 | |
| Certificate of Analysis | Apr 27, 2023 | T494067 | |
| Certificate of Analysis | Apr 27, 2023 | T494067 | |
| Certificate of Analysis | Apr 27, 2023 | T494067 |