A colorimetric peroxidase substrate
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Alternative(s)
36361TMB
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TMB (hydrochloride hydrate)

Item No. 70450

Product Insert (PDF)Safety Data Sheet (SDS) (PDF)
Technical Information
Formal Name
3,3',5,5'-tetramethyl-[1,1'-biphenyl]-4,4'-diamine, dihydrochloride, hydrate
CAS Number
207738-08-7
Synonyms
  • 3,3′,5,5′-Tetramethylbenzidine
Molecular Formula
C16H20N2 • 2HCl [XH2O]
Formula Weight
313.2
Purity
≥98%
Formulation (Request formulation change)
A crystalline solid
Solubility(Learn about Variance in Solubility)
DMSO: 1 mg/mlDMSO:PBS (1:300): 30 µg/ml
λmax
295 nm
SMILES
CC1=C(N)C(C)=CC(C2=CC(C)=C(N)C(C)=C2)=C1.Cl.Cl.O
InChi Code
InChI=1S/C16H20N2.2ClH.H2O/c1-9-5-13(6-10(2)15(9)17)14-7-11(3)16(18)12(4)8-14;;;/h5-8H,17-18H2,1-4H3;2*1H;1H2
InChi Key
KVCWTKDFVVSVSJ-UHFFFAOYSA-N
Shipping & Storage Information
Storage
-20°C
Shipping
Room temperature in continental US; may vary elsewhere
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    100 mg

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    1 g

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    Product Description

    TMB is a colorimetric substrate for peroxidases, including hemoglobin, myeloperoxidase, and horseradish peroxidase (HRP).1,2,3,4 Enzymatic oxidation of TMB produces a radical cation followed by a diimine product that can be quantified by colorimetric detection at 652 and 450 nm, respectively, as a measure of peroxidase activity. TMB has commonly been used as a substrate for HRP in ELISA and immunohistochemical applications.5,6

    WARNING This product is not for human or veterinary use.

    References & Product Citations
    Product Description References

    1. Thomas, P.D., and Poznansky, M.J. A modified tetramethylbenzidine method for measuring lipid hydroperoxides. Anal. Biochem. 188(1), 228-232 (1990).

    2. Marquez, L.A., and Dunford, H.B. Mechanism of the oxidation of 3,5,3',5'-tetramethylbenzidine by myeloperxidase determined by transient-and steady-state kinetics. Biochemistry 36(31), 9349-9355 (1997).

    3. Josephy, P.D., Eling, T., and Mason, R.P. The horseradish peroxidase-catalyzed oxidation of 3,5,3',5'-tetramethylbenzidine. Free radical and charge-transfer complex intermediates. The Journal of Biological Chemisty 257(7), 3669-3675 (1982).

    4. Madersbacher, S., and Berger, P. Double wavelength measurement of 3,3',5,5'-tetramethylbenzidine (TMB) provides a three-fold enhancement of the ELISA measuring range. J. Immunol. Methods 138(1), 121-124 (1991).

    5. Reyna-Bello, A., Eleizalde, M.C., and Silva, A.M. Assessment of chromogen suitability in ELISA for the detection of anaplasmosis and trypanosomosis. J. Immunoassay Immunochem. 28(1), 1-11 (2007).

    6. Mesulam, M.M., and Rosene, D.L. Sensitivity in horseradish peroxidase neurohistochemistry: A comparative and quantitative study of nine methods. J. Histochem. Cytochem. 27(3), 763-773 (1979).