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3,3,5,5-Tetramethylbenzidine, 1 g, glass

3,3,5,5-Tetramethylbenzidine, 1 g, glass

SKU 6350.1
€ 69,60
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3,3,5,5-Tetramethylbenzidine, 1 g, glass
Product Details
CAS number: 9048-46-8
HS code: 29215990
Brand: Carl Roth
3,3',5,5'-Tetramethylbenzidine, min. 98 %, p.a., Molar mass (M) 240,35 g/mol, Storage temp. +4 °C, CAS No. [54827-17-7], EG-Nr. 259-364-6, Empirical formula C16H20N2


3,3',5,5'-Tetramethylbenzidine ≥98 %, p.a.

Used as a highly sensitive substrate for peroxidases, particularly in ELISA procedures. Can replace carcinogenic benzidine.


Dissolves easily in acetone, chloroform and ethanol.


Gebruiksinstructies

Working solution: 1 mg TMB in 0.1 ml dimethyl sulfoxide, add 9.9 ml 0.1 M sodium acetate (pH 6). Filter and add H2O2 ad 0.01 %. Always prepare freshly!
Application: approx. 50 µl per 96well, incubate for 10-30 mins. at room temperature. Add 50 µl 1 M H2SO4 and quantitate photometrically at 450 nm (acc. to: Bos et. al. (1981) J. Immunoassay 2:187).
Protect from light.



Technische informatie
Enzyme Horseradish Peroxidase
Bewijs Colour (blue)
Signaalproduct soluble (ELISA)
Assay protocol for use of BCIP/NBT in immunoblot procedures:

Stock solutions (All solutions are stable for at least 1 year at 4 °C):
0.5 g NBT in 10 ml 70 % dimethylformamide;
0.5 g BCIP (p-toluidine salt) in 10 ml 100 % dimethylformamide.
Incubation buffer for alkaline phosphatase:
100 mM NaCl, 5 mM MgCl2,100 mM Tris (pH 9.5).
Fresh substrate solution: 66 μl NBT stock solution + 10 ml incubation buffer, mix well, add 33 μl BCIP stock solution. Use within 1 hour.
Blot development: Approx. 10 ml substrate solution per 15 x 15 cm2 membrane surface. Develop at room temperature until bands become visible (approx. 30 mins).
Reaction stop: Rinse with PBS/20 mM EDTA.


Detection of peroxidase activity (Immunoassay):

Dissolve 1 mg TMB in 0.1 ml dimethylsulfoxide (Art. No. 4720); add 9.9 ml of a 0.1 M sodium acetate solution (pH 6.0) (Powder: Art. No. 6779), filter and add H2O2 (Art. No. 8070) (final concentration 0.01 %).

Always prepare freshly!

Incubation 10-30 mins at room temperature (approx. 50 μl per microtitre well; finally add 50 μl 1 M H2SO4, Art. No. X873, per well).
Photometric quantitation at 450 nm.

Reference: Bos E.S. et al. (1981) J. Immunoassay 2, (3/4), 187.


Appearance beige cryst. powder
Assay (GC) ≥99.0 %
Sulphated ash ≤0,2 %
Loss on drying (3 h, 60 °C, vacuum) ≤1,0 %
Melting range 166-171 °C
Can replace carcinogenic benzidine.
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