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TMB Stabilized Substrate for Horseradish Peroxidase

by Promega
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  • Overview
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  • TMB Stabilized Substrate is a stable, ready-to-use TMB (3,3´, 5,5´-tetramethylbenzidine) color development substrate for localization of horseradish peroxidase-conjugated antibodies on dot blots and Western blots. It is easier to use than 4-chloro-1-naphthol (CN), which must be prepared immediately before use. TMB Stabilized Substrate comes premixed and fully diluted in a proprietary buffer containing less than 0.5% organic solvent.

    TMB Stabilized Substrate is at least threefold more sensitive than 4-chloro-1-naphthol (CN); as little as 412pg of ?-galactosidase detected on TMB blot as compared to 1.12ng on CN blot when detected with a ?-galactosidase-specific antibody and HRP-conjugated secondary antibody. The color is much more stable than 4-chloro-1-naphthol and photographs more easily.

    Note: Not recommended for use with ELISAs.

    1. Larson, G. (1992) Promega Notes 39, 17–18.
    2. Cullinane, A. et al. (2007) Diagnosis of equine infectious anaemia during the 2006 outbreak in Ireland. Vet. Rec. 161, 647–52.
    3. Tetteh, K.K.A. et al. (2005) Extensive antigenic polymorphism within the repeat sequence of the Plasmodium falciparum merozoite surface protein 1 block 2 is incorporated in a minimal polyvalent immunogen. Infect. Immun. 73, 5928–35.
    4. Stenbak, C.R. and Linial, M.L. (2004) Role of the C terminus of foamy virus Gag in RNA packaging and pol expression. J. Virol. 78, 9423–30.
    5. Mello, K. et al. (2002) A multigene family that interacts with the amino terminus of plasmodium MSP-1 identified using the yeast two-hybrid system. Eukaryot. Cell 1, 915–25.
    Western blot (immunoblot) for ?-actin in cytoplasmic lysate from HEK293T cells.
    Western blot (immunoblot) for ?-actin in cytoplasmic lysate from HEK293T cells.

    Blot performed on a serial dilution of lysate; each lane contains an indicated amount of total protein (µg). Primary antibody (monoclonal) used at 1:5,000; Anti-Mouse IgG, HRP Conjugate secondary antibody (Cat.# W4021) used at 1:2,500; blot imaged with TMB Stabilized Substrate for Horseradish Peroxidase (Cat.# W4121).

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    Storage Conditions

    +15C TO +30C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.