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TMB Stable Liquid Substrate 1.25mM for HRPO (3,3',5,5'-Tetramethylbenzidine)

Cat no: T6005

TMB Stable Liquid Substrate 1.25mM for HRPO (3,3',5,5'-Tetramethylbenzidine)

Tetramethylbenzidine (TMB) is an excellent substrate for detecting horseradish peroxidase (HRP) labeled probes. TMB produces a soluble end product which is blue in color that is quantitated at 650nm, providing an ultrasensitive quantitative substrate system. \n\nTMB has been shown to be a safe-sensitive substrate for the assay of horseradish peroxidase (HRP). Initially, in the presence of HRP and hydrogen peroxide, a one-electron oxidation product is formed.\n\nThis compound, a cation free radical, is blue in color wth an adsorption maximum at 653nm. Further reaction with HRP/H2O2 or acidification of the radical with acid yields the diimine terminal oxidation product adsorbing light at 450 nm. The extinction coefficient of the radical (E653 nm = 5.9x10e4 mol-1 cm-1) provide a remarkably sensitive system for the assay of HRP and HRP labeled probes.\n\nTMB is a single component reagent stable at room temperature and not sensitive to normal laboratory light. It is optimized with respect to TMB and hydrogen peroxide concentrations and yields a linear response with the concentrations of HRP usually employed in immunologic assays.\n\nTMB reagents for ELISA and blotting techniques have been available for several years. Histochemically, TMB has been utilized primarily for the study of retrograde axonal transport. To prevent needle formation during color development, sodium nitroprusside in combination with a low pH was used.\n\nRecent publications have described stabilization of the TMB precipitate with ammonium hexamolybdate or sodium tungstate. Methyl salicylate has also been employed to render the TMB reaction product insoluble in alcohols.\n\nUnited States Biological supplies a stable, single component TMB for histochemical use containing none of these stabilizers and enhancers. The final reaction product is not affected by ethanol, xylene or xylene substitutes. Thus dehydration through alcohols to xylene and mounting the sections in a xylene type medium is possible.\n\nTMB may also be used for blotting procedures.

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SPECIFICATIONS

Catalog Number

T6005

Size

250ml

Applications

ELISA, WB

References

US Biological application reference: Poulain-Godefroy, O. and Froguel, P., BBRC 356: 662-667 (2007).

Alternative Names

3,3

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