TMB Peroxidase Substrate

TMB (tetramethylbenzidine) peroxidase substrates are chromogenic substrates used in ELISA applications. In a chromogenic or colorimetric based ELISA, HRP (horseradish peroxidase) is used as a common detection enzyme in the development of immunoassays for diagnostic applications is horseradish peroxidase (HRP), which is a small, stable enzyme, ~40 kDa, that can be easily conjugated to an antibody, hapten, or peptide. Several substrates are available for use with HRP, but the most common is TMB. Surmodics™ IVD offers several choices for one component TMB substrates with varying kinetic rates for optimization of your assays.

Surmodics IVD’s TMB peroxidase substrate solution products provide unparalleled and consistent performance for ELISA applications. Our BioFX™ TMB peroxidase substrates offer assay developers best in class sensitivity, stability and low background in a one-component format.

Surmodics IVD’s TMB peroxidase substrates are all offered in ready-to-use formulations that incorporate easily into the existing assay manufacturing process with varying kinetic rates for optimization of your assays.

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TMB Peroxidase Microwell Substrates

Surmodics IVD is a leading provider of TMB peroxidase microwell substrates. These reagents are the gold standard for lot-to-lot consistency, sensitivity, stability and overall help assay developers to achieve their optimum level of detection. There are several technical advantages when using our TMB peroxidase substrates including the following:

  • Best-in-Class Kinetic Range:
    • Surmodics offers assay developers a wide range of kinetic rates for their unique assay needs.
  • Outstanding Signal Generation:
    • BioFX TMB substrates generate higher signal per femtogram of analyte versus other TMB substrates.
  • Reliable Durability:
    • Surmodics’ TMB substrates maintain robust formulation stability.
  • Ease of Manufacture
    • ISO 13485:2016 & 9001:2015 certified.
    • Expect lot-to-lot consistency, reproducibility, and optimal performance.

Surmodics IVD’s TMB Peroxidase Microwell Substrates:

  • NEW BioFX High Definition One Component HRP Microwell Substrate
    • Product Code: TMBHD
    • Gives assay developers maximal signal generation and low backgrounds while maintaining a long shelf life.
  • BioFX TMB Conductivity One Component HRP Microwell Substrate
    • Product Code: TMBC
    • Mid-kinetic range substrate for ELISA applications.
  • BioFX TMB Super Sensitive One Component HRP Microwell Substrate
    • Product Code: TMBS
    • Gives assay developers maximal signal generation and low backgrounds while maintaining a long shelf life.
  • TMB One Component HRP Microwell Substrate
    • Product Code: TMBW
    • Gives assay developers the best possible tool to maximize shelf life stability and lower backgrounds while setting the industry standard for signal generation of the standard curve.
  • TMB Extended Range HRP Microwell Substrate
    • Product Code: TMBX
    • Low-kinetic range substrate for ELISA applications.
  • TMB Double Slow One Component HRP Microwell Substrate
    • Product Code: TMDS
    • low-kinetic range substrate for ELISA applications.
  • TMB Slow Kinetic One Component HRP Microwell Substrate
    • Product Code: TMSK
    • mid-kinetic range substrate for ELISA applications.
  • TMB Super Slow One Component HRP Microwell Substrate
    • Product Code: TTMB
    • low-kinetic range substrate for ELISA applications.
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TMB Peroxidase Membrane/Precipitating Substrates:

In addition to our TMB peroxidase microwell substrates, Surmodics IVD also offers TMB peroxidase membrane substrates. Our TMB peroxidase membrane substrates outperform competitors for low background, stability and sensitivity.

Surmodics offers both HRP and AP formulations of membrane substrates to provide a “one stop shop” fora variety of applications including the development of point-of-care technologies, dot blots, western blots, ELISpots and more.

Our TMB peroxidase membrane substrates also offer unparalleled consistency & quality. Along with all Surmodics IVD’s reagents, they are manufactured under stringent conditions that minimize variation among production lots. Surmodics is an ISO 13485/9001 certified and a 21CFR820 compliant facility and is committed to delivering quality products.

Surmodics IVD’s TMB Peroxidase Membrane/Precipitating Substrates:

  • TMB Enhanced HRP Membrane Substrate
    • Product Code: ESPM
    • Provides an intense staining pattern for increased sensitivity.
    • Surmodics’ TMB Enhanced HRP Membrane Substrate (ESPM) provides extended dynamic range with reduction of wash out at higher peroxidase concentrations. ESPM demonstrates similar sensitivity to most mid-to-high level chemiluminescent substrates without the costly equipment needed for detection.
  • TMB One Component HRP Membrane Substrate
    • Product Code: TMBM
    • Provides increased sensitivity with low backgrounds.
    • TMB One Component HRP Membrane Substrate (TMBM) staining pattern provides a dark blue precipitate with low background to increase the signal-to-noise ratios in western blot, dot blots and line assays.

Visit the link below to request free samples of our TMB peroxidase substrates.

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Criteria for Choosing TMB Peroxidase Substrates:

During assay development it is important to consider your substrate selection carefully and to have an understanding of your assay’s requirements. Several key elements can be important such as dynamic range, kinetic rate, and assay timing.

Dynamic range is especially important when developing an assay that requires quantitation of a large range of analyte concentrations. Kinetic rate is also an important criterion when finding the right substrate for your ELISA. As such, Surmodics IVD offers several TMB peroxidase substrates at varying kinetic rates to ensure we have an option suitable for each unique assay in question.

In addition, the timing of your assay is important when considering factors such as dynamic range, reproducibility and detection limit. Assay timing can be optimized by choosing a TMB peroxidase substrate with the kinetic rate that allows your assay to be performed within the desired timeframe.

Key assay parameters, such as dynamic range, kinetic rate and assay timing are important factors to consider when evaluating various TMB peroxidase substrates. Each TMB peroxidase substrate impacts these parameters differently. Choosing the best substrate for your assay will allow the assay developer to create a robust assay with superior performance.

Intended Use of TMB Peroxidase Substrate

Due to the unique way the TMB Peroxidase substrate acts when it interacts with enzymes, it is possible to use this substrate product for a wide variety of applications including ELISA procedures. It can cause the solution to turn a faint blue color. This specific hue of blue allows assay developers and researchers to utilize the TMB peroxidase substrate to determine specific bioassay characteristics. The TMB peroxidase substrate allows it to donate hydrogen during the reduction of peroxidase substrate enzymes. The TMB peroxidase substrate changes to absorb a wavelength of light of a known value that can be readily measured by aiming a spectrophotometer at the substrate. If the product or the substrate is exposed to acidic conditions the solution will turn a yellow color instead which can also be measured easily with a spectrophotometer.

Applications of TMB Peroxidase Substrate

The TMB substrate product is recommended to use with ELISA peroxidase-based enzyme immunoassays. The product can be used by adding the TMB substrate to the peroxidase reaction. If the substrate is measured carefully and is able to react as with a peroxidase it will be able to donate a hydrogen. Whether the substrate donates a hydrogen to the peroxidase enzyme also depends on if the peroxidase is in a basic or acidic solution.