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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. This substrate gives higher signal and has a higher sensitivity in ELISA than OPD (ortho-phenylenediamine dichloride)
TMB X-tra characteristics are:
High sensitivity
Stable and robust
Aqueous based – no organic solvents (REACH Annex XVII compliant)
Low lot-to-lot variation
For traceable pipetting, we recommend using pink colored TMB X-tra Prestained (cat. no. T494156-01). The pink indicator disappears after addition of the stop solution.
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 X-tra is a ready-to-use, very active and stable aqueous formulation. The substrate gives a very high signal for a particularly fast assay reaction. The composition of the product makes it very resistant to raised temperatures. 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 100µL 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 unless a corresponding TMB X-tra Diluent is acquired. The diluted TMB solution maintains the same stability performance as the undiluted version, and a specific activity can be achieved.
• For traceable pipetting, we recommend using the pink colored TMB X-tra Prestained (cat. no. T494156-01). The pink indicator disappears after addition of the stop solution.
• In case of incidents of precipitation with very high OD-values, we 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.
Product specifications and parameters
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Comprehensive hazard, handling, storage, and regulatory compliance document.
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| Lot Number | Certificate Type | Data | Oggetto |
|---|---|---|---|
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 | |
| Certificate of Analysis | Apr 27, 2023 | T494156 |