TMB X-tra - Ready-to-use (RTU), high purity

Item Number

T494156

• Synonyms: TMB
• Specifications & Purity: Ready-to-use (RTU)
• Storage Temp: Store at 2-8°C,Protected from light
• Shipped In: Wet ice

Product Description

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

SKU	Product name	Product Type	Pack Size
T494156-01-100ml	TMB X-tra	Prestained	100ml
T494156-01-500ml	TMB X-tra	Prestained	500ml
T494156-01-1L	TMB X-tra	Prestained	1L
T494156-02-100ml	TMB X-tra	Not prestained	100ml
T494156-02-500ml	TMB X-tra	Not prestained	500ml
T494156-02-1L	TMB X-tra	Not prestained	1L