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Soluble TMB Substrates


Slow, fast and ultrasensitive varieties of the most popular ELISA substrate.

Pierce TMB Substrates for ELISA detection HRP conjugates.

Thermo Scientific Pierce TMB ELISA Substrates detect horseradish peroxidase (HRP) activity, yielding a blue color (Amax = 370nm and 652nm) that changes to yellow (Amax = 450nm) upon addition of a sulfuric or phosphoric acid stop solution.

TMB is the most popular chromogenic substrate for HRP detection in ELISA and is available in several formats. The Pierce 1-Step TMB Substrates are one-component substrates that require no preparation before use. Unlike other commercially available substrates, these products contain no DMF or DMSO.

Highlights:

  • Available in traditional or ready-to-use formulations
  • No additional reagents or filtering required
  • Non carcinogenic
  • Various levels of sensitivity and development speed to suit any assay
  • No DMF or DMSO present in the reagents

Product Details:

Comparison of Thermo Scientific ELISA Substrates.
Comparison of sensitivities of Thermo Scientific TMB ELISA Substrates.

Thermo Scientific Pierce 1-Step Ultra TMB yields the greatest sensitivity among the TMB substrates, followed by 1-Step Turbo TMB and 1-Step Slow TMB. The sensitivity of the 1-Step Turbo TMB is similar to OPD-based substrates formulated at approximately 1mg/mL. 1-Step Slow TMB has intermediate sensitivity – more sensitive than ABTS, but less sensitive than o-phenylenediamine dihydrochloride (OPD) or 1-Step Turbo TMB. The 1-Step Slow TMB is an ideal substrate for kinetic studies.

Comparison of sensitivities of Thermo Scientific 1-Step Turbo TMB, Slow TMB and ABTS ELISA Substrates. Comparison of sensitivities of Thermo Scientific 1-Step Turbo TMB, Slow TMB and ABTS ELISA Substrates.

TMB (3,3′,5,5′-tetramethylbenzidine) is a chromogen that yields a blue color when oxidized, typically as a result of oxygen radicals produced by the hydrolysis of hydrogen peroxide by HRP. For kinetic or non-stopped ELISA assays, the TMB chromogen has maximal absorbances at 370nm and 652nm. The color then changes to yellow with the addition of sulfuric or phosphoric acid with maximum absorbance at 450nm. A green reaction product may result from partial conversion to the yellow product from the blue intermediate. TMB is very sensitive ELISA substrate and is more quickly oxidized than other HRP substrates, resulting in faster color development.

Thermo Scientific 1-Step Ultra TMB-ELISA Substrate produces more signal than competing products. Thermo Scientific 1-Step Ultra TMB-ELISA Substrate produces more signal than competing products.

Thermo Scientific 1-Step Ultra TMB-ELISA Substrate provides better signal-to-noise ratio than competing products. Thermo Scientific 1-Step Ultra TMB-ELISA Substrate provides better signal-to-noise ratio than competing products.

References:

  1. Josephy, P.D., et al. (1982). J. Biol. Chem. 257(7), 3669-3675.
  2. Josephy, P.D., et al. (1983). J. Biol. Chem. 258(9), 5561-5569.
  3. Liem, H.H., et al. (1979). Anal. Biochem. 98, 388-393.
  4. Thomas, P.E., et al. (1976). Anal. Biochem. 75, 168-176.
  5. Hong, P.W., et al. (2002). J. Virol. 76, 12855-12865.
  6. Tek, V. and Zolkiewski, M. (2002). Protein Sci. 11, 1192-1198.
  7. Wu, S.-C. and Wong, S.-L. (2002). Appl. Envir. Microbiol. 68, 1102-1108.
  8. Su, S.V., et al. (2004). J. Biol. Chem. 279(18), 19122-19132.
  9. Weimer, B.C., et al. (2001). Appl. Envir. Microbiol. 67, 1300-1307.
  10. Murphy, M.B., et al. (2003). Nucleic Acids Res. 31, e110.

Related Resources:

Assay development and ELISA technical guide
ELISA substrate selection guide
Spike-and-recovery and linearity-of-dilution assessment

Related Products:

Precipitating TMB Substrates for Western blotting
ABTS
OPD (o-Phenylenediamine)
PNPP Substrates
Coated and Uncoated Microwell Plates
ELISA Products


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
34021 TMB Substrate Kit
Formulation: Two-component kit containing 3,3',5,5'-tetramethylbenzidine (0.4g/L) and peroxide (0.02% H202) solutions
Sufficient For: 400mL of typical 1X TMB buffer (4000 wells at 100μL per well)

Kit Contents:
TMB Solution, 200mL
Peroxide Solution, 200mL
400mL kit Product Instructions for product #34021 TMB Substrate Kit MSDS for product #34021 TMB Substrate Kit Certificate of Analysis for product #34021 TMB Substrate Kit $134.00

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34022 1-Step Turbo TMB-ELISA Substrate Solution
Formulation: Ready-to-use (1X) buffered solution of TMB and peroxide
Sufficient For: 2500 wells at 100μL per well
250mL Product Instructions for product #34022 1-Step Turbo TMB-ELISA Substrate Solution MSDS for product #34022 1-Step Turbo TMB-ELISA Substrate Solution Certificate of Analysis for product #34022 1-Step Turbo TMB-ELISA Substrate Solution $121.00

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34024 1-Step Slow TMB-ELISA Substrate Solution
Formulation: Ready-to-use (1X) buffered solution of TMB and peroxide
Sufficient For: 2500 wells at 100μL per well
250mL Product Instructions for product #34024 1-Step Slow TMB-ELISA Substrate Solution MSDS for product #34024 1-Step Slow TMB-ELISA Substrate Solution Certificate of Analysis for product #34024 1-Step Slow TMB-ELISA Substrate Solution $88.00

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34028 1-Step Ultra TMB-ELISA Substrate Solution
Formulation: Ready-to-use (1X) buffered solution of TMB and peroxide
Sufficient For: 2500 wells at 100μL per well
250mL Product Instructions for product #34028 1-Step Ultra TMB-ELISA Substrate Solution MSDS for product #34028 1-Step Ultra TMB-ELISA Substrate Solution Certificate of Analysis for product #34028 1-Step Ultra TMB-ELISA Substrate Solution $136.00

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34029 1-Step Ultra TMB-ELISA Substrate Solution
Formulation: Ready-to-use (1X) buffered solution of TMB and peroxide
Sufficient For: 10000 wells at 100μL per well
1L Product Instructions for product #34029 1-Step Ultra TMB-ELISA Substrate Solution MSDS for product #34029 1-Step Ultra TMB-ELISA Substrate Solution Certificate of Analysis for product #34029 1-Step Ultra TMB-ELISA Substrate Solution $440.00

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34028B 1-Step Ultra TMB - ELISA Substrate Custom MSDS for product #34028B 1-Step Ultra TMB - ELISA Substrate Certificate of Analysis for product #34028B 1-Step Ultra TMB - ELISA Substrate

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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