Application :

Suitable for the enzyme immuno assay of horseradish peroxidase (HRP) & molecular biology.

3,3,5,5,Tetramethylbenzidine (TMB) is a colorimetric substrate used with peroxidase and peroxidase-coupled systems. It produces a soluble end product, which is blue in color and can be read spectrophotometrically at 370 nm or 655 nm( photometric mode). The reaction can be stopped with acid (Trichloroacetic acid or Sulphuric Acid or as per application). This product is also been used for hematology and immunochemical tests.

Preparation Instructions:

A TMB stock solution can be prepared by dissolving 10 mg/ml in DMSO(41.6 mM sol). The working solution is made by diluting 0.1 ml of the DMSO stock with 9.9 ml Sodium Citrate buffer, pH 6.0,(sodium Acetate (0.7M) and adjust ph by using citric Acid 6.0) and then adding 2ul of fresh 30% hydrogen peroxide per 10 ml of substrate/buffer solution immediately prior to use. The hydrochloride salt can be directly dissolved in aqueous media. For this reason, the hydrochloride salt is the preferred form of TMB; however, both forms will give identical results.

Stock solutions of TMB in DMSO can be frozen in 1–2 ml aliquots at –20deg C. Solutions may appear to remain active for at least one years when stored in this manner. Any oxidation would be revealed as a color change in the working solution.

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