3,3',5,5'-TETRAMETHYLBENZIDINE DIHYDROCHLORIDE HYDRATE Chemical Properties

Melting point:

>300°C

storage temp. 

2-8°C

solubility 

Soluble in DMSO (~1.7 mg/ml), and 1:300 DMSO:PBS (pH 7.2) (0.03 mg/ml).

form 

Powder

color 

Light brown to brown

Sensitive 

Moisture & Light Sensitive

InChI

InChI=1S/C16H20N2.ClH.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;1H;1H2

InChIKey

NYNRGZULARUZCC-UHFFFAOYSA-N

SMILES

CC1=C(C(C)=CC(C2C=C(C)C(N)=C(C)C=2)=C1)N.Cl.O

CAS DataBase Reference

207738-08-7

Safety Information

Risk Statements 

22

Safety Statements 

22-24/25-25-24

WGK Germany 

3

TSCA 

Yes

HS Code 

29215900

3,3',5,5'-TETRAMETHYLBENZIDINE DIHYDROCHLORIDE HYDRATE Usage And Synthesis

Description

TMB is an aromatic amine that undergoes oxidation by the higher oxidation states of heme peroxidases (compounds I and II) thereby serving as a reducing co-substrate. One electron oxidation of TMB results in a radical cation that forms a charge-transfer complex with the unoxidized compound. This charge transfer complex absorbs at 652 nm (ε = 39,000). The completely oxidized form (diimine) absorbs at 450 nm (ε = 59,000) and is formed by two sequential one-electron oxidations of TMB. Thus the stoichiometry of oxidation is 0.5 mole charge transfer complex (λ_max = 652 nm) or 1 mole of diimine (λ_max = 450 nm) formed (or TMB oxidized) per mole of hydroperoxide reduced by the peroxidase.

Chemical Properties

Off-white fluffy powder

Uses

3,3',5,5'-Tetramethylbenzidine dihydrochloride hydrate is a chromophore substrate in ELISA assays. It is a noncarcinogenic substitute for benzidine.

Uses

Convenient water-soluble form of TMB for determination of peroxidase.

Uses

3,3′,5,5′-Tetramethylbenzidine dihydrochloride hydrate has also been used as a horseradish peroxidase substrate in enzyme linked immunosorbent assay and reverse dot blot (RDB).

General Description

3,3′,5,5′-Tetramethylbenzidine dihydrochloride hydrate (TMB-d) is a sensitive chromogen substrate for peroxidase. It is non- carcinogenic, hence, it can be used as a safe substitute for benzidine. TMB-d is useful in quantifying hemoglobin and in staining myeloperoxidase-containing granules in granulocytes.

References

[1] PANAKKEZHUM D. THOMAS  Mark J P. A modified tetramethylbenzidine method for measuring lipid hydroperoxides[J]. Analytical biochemistry, 1990, 188 1: Pages 228-232. DOI: 10.1016/0003-2697(90)90557-p
[2] LEAH A. MARQUEZ  H. B D. Mechanism of the Oxidation of 3,5,3‘,5‘-Tetramethylbenzidine by Myeloperoxidase Determined by Transient- and Steady-State Kinetics?[J]. Biochemistry Biochemistry, 1997, 36 31: 9349-9355. DOI: 10.1021/bi970595j
[3] P. JOSEPHY R M T Eling. The horseradish peroxidase-catalyzed oxidation of 3,5,3’,5’-tetramethylbenzidine. Free radical and charge-transfer complex intermediates.[J]. The Journal of Biological Chemistry, 1982, 31 1: 3669-3675. DOI: 10.1016/s0021-9258(18)34832-4
[4] S. MADERSBACHER  P. B. Double wavelength measurement of 3,3′,5,5′-tetramethylbenzidine (TMB) provides a three-fold enhancement of the ELISA measuring range[J]. Journal of immunological methods, 1991, 138 1: Pages 121-124. DOI: 10.1016/0022-1759(91)90071-m
[5] ARMANDO REYNA-BELLO  Adriana M S  Mariana C Eleizalde. Assessment of chromogen suitability in ELISA for the detection of anaplasmosis and trypanosomosis.[J]. Journal of immunoassay & immunochemistry, 2007, 28 1: 1-11. DOI: 10.1080/15321810601025432

3,3',5,5'-TETRAMETHYLBENZIDINE DIHYDROCHLORIDE HYDRATE(207738-08-7)Related Product Information

• 4-AMINO-3-METHYLBIPHENYL
• 3,3',5,5'-Tetramethylbenzidine dihydrochloride
• 2,6-DIMETHYLANILINE HYDROCHLORIDE
• O-Tolidine
• 3'-METHYL-BIPHENYL-4-YLAMINE
• Tetramethylbenzidine
• 3,3',5,5'-TETRAMETHYLBIPHENYL
• 4-Aminobiphenyl hydrochloride
• Benzidine dihydrochloride
• 2,4,6-Trimethylaniline
• 3,3'-Dimethylbenzidine dihydrochloride
• 2,4-Dimethylaniline hydrochloride
• 3,3'-Dimethylbiphenyl
• 3,5-DIMETHYL-BIPHENYL
• 3,3',5,5'-TETRAMETHYLBENZIDINE DIHYDROCHLORIDE HYDRATE