Name: TMBZ HCl
Price: 394.00 USD

- Overview
  - TMBZ is a chromogenic reagent utilized for peroxidase detection. It has been developed as an alternative to benzidine, which is a carcinogenic chemical. Because of the ortho methyl groups on its benzene ring, TMBZ is not metabolized into highly carcinogenic o-hydroxybenzidines or o,o'- dihydroxybenzidines. Therefore, TMBZ compounds are much less carcinogenic than benzidine. Although the TMBZ solution is colorless, it turns bluish-green (λmax: 655 nm) in the presence of hydrogen peroxide and peroxidase. The structure of this bluish-green complex is thought to be a radical form of two oxidized TMBZ molecules. TMBZ HCl is a hydrochloride form of TMBZ that is readily soluble in water (100 mg TMBZ HCl per 10 ml water).
    
    Please contact us at for specific academic pricing.
- Properties
  - Categories
    
    Oxidative Chromogenic Dyes
    
    CAS Number
    
    64285-73-0
    
    Molecular Formula
    
    C16H22Cl2N2· 2H2O
    
    Molecular Weight
    
    349.3
    
    Appearance
    
    white or pale yellow powder
    
    Purity
    
    >98.0% (Titration)
    
    Storage
    
    ambient temperature
    
    Shipping
    
    ambient temperature
    
    * For research use only
- Applications
  - Application Description
    
    Assay Protocol
    1. Dissolve 6 mg of TMBZ with 1 ml DMSO to prepare 100X TMBZ solution.
    2. Mix 5 μl of 30% hydrogen peroxide solution with 1 ml PBS to prepare 200X H2O2 solution.
    3. Add 10 μl of 100X TMBZ solution and 5 μl of 200X H2O2 solution to 1 ml PBS to prepare staining solution.*
    4. Add 100-200 μl of the staining solution to each well of a 96-well microplate. Incubate the plate at room temperature or at 37°C for 5 minutes to 1 hour.
    5. Wash the sample with PBS several times to stop the staining reaction.
    * For the best results, modification of the final concentration of TMBZ and hydrogen peroxide may be necessary. The staining solution is not stable. Please prepare fresh solution prior to use.
- Reference
  - 1) V. R. Holland, et al., A Safer Substitute for Benzidine in the Detection of Blood. Tetrahedron. 1974;30:3299-3302.
    
    2) W. Levin, et al., Preparation of Partially Purified, Lipid-depleted Cytochrome P-450 and Reduced Nicotinamide Adenine Dinucleotide Phosphatecytochrome c Reductase from Rat Liver Microsomes. J Biol Chem. 1974;249:1747-1754.
    
    3) P. E. Thomas, et al., An Improved Staining Procedure for the Detection of the Peroxidase Activity of Cytochrome p-450 on Sodium Dodecyl Sulfate Polyacrylamide Gels. Anal Biochem. 1976;75:168-176.
    
    4) H. H. Liem, et al., Quantitative Determination of Hemoglobin and Cytochemical Staining for Peroxidase Using 3, 3'- 5, 5'-Tetramethylbenzidine dihydrochloride, a Safe Substitute for Benzidine. Anal Biochem. 1979;98:388-393.
    
    5) R. C. Lijana, et al., Tetramethylbenzidine- A substitute for Benzidine in Hemoglobin Analysis. J Lab Clin Med. 1979;94:266-276.
    
    6) R. M. Jaffe, et al., A New Occult Blood Test Not Subject to False-Negative Results from Reducing Substances. J Lab Clin Med. 1979;93:879-886.
    
    7) K. Suzuki, et al., Assay Method for Myeloperoxidase in Human Polymorphonuclear Leukocytes. Anal Biochem. 1983;132:345-352.
    
    8) F. H. Pujol, et al., A Double Sandwich Monoclonal Enzyme Immunoassay for Detection of Hepatitis B Surface Antigen. J Immunoassay. 1993;14:21-31.
    
    9) F. B. Serrat, Colorimetric Method for Determiantion of Chlorine with 3, 3'- 5, 5'-Tetramethylbenzidine. Talanta. 1994;41:2091-2094.

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