Activity of Neogen's TMB Substrates

Neogen uses K-Blue substrates to manufacture thousands of assays each year for a range of applications. Key areas of Neogen’s substrate use include assays for forensic toxicology, therapeutic drug monitoring, food safety testing, and life science research.

As an immunoassay manufacturer, Neogen recognises that one TMB (3,3',5,5'-tetramethylbenzidine) microwell substrate formulation will not meet the specifications of all HRP-based (horseradish peroxidase) immunoassays. Therefore, Neogen offers multiple unique one-bottle TMB microwell colourimetric substrate formulations to meet specific requirements for different assay systems.

Neogen’s TMB microwell chromogenic substrate formulations are used with horseradish peroxidase (HRP) immunoassays. Our TMB substrates turn a deep blue colour in the presence of horseradish peroxidase labelled conjugate. These substrates are not applicable for use with assays requiring a precipitating substrate. Neogen substrates have common characteristics of low background, excellent lot-to-lot consistency and long term stability. Each formula offers sensitive peroxidase enzyme detection and unique performance characteristics allowing assay manufacturers to select the perfect chromogenic solution for their HRP assay requirements.

Request Substrate

Available Products

Microwell HRP Substrates

ProductActivityShelf LifeProduct FeaturesAvailable Sizes
K-Blue Advanced Highest activity  24 months when stored at 2-8°C
  • Superior activity
  • RTU (ready-to-use)
  • Low background
  • High consistency between lots
  • 200 mL
  • 500 mL
  • 1 L
  • 20 L (1 X 20L)
Enhanced K-Blue High activity

48 months when stored at 2-8°C
  • Longest shelf-life of Neogen TMB substrates
  • RTU (ready-to-use)
  • Low background
  • High consistency between lots
  • 200 mL
  • 500 mL
  • 1 L
  • 20 L (1 x 20 L)
Custom TMB Reduced activity: 20–30% lower activity than Enhanced K-Blue

36 months when stored at 2-8°C
  • Ideal for assays requiring a less active substrate
  • RTU (ready-to-use)
  • Low background
  • High consistency between lots
  • 200 mL
  • 500 mL
  • 1 L
  • 20 L (1 x 20 L)
K-Blue Aqueous Mid-level activity 36 months when stored at 2-8°C
  • 100% solvent free
  • Ideal when working under strict regulatory requirements
  • RTU (ready-to-use)
  • Low background
  • High consistency between lots
  • 200 mL
  • 500 mL
  • 1 L
  • 20 L (1 x 20 L)
Alert K-Blue Aqueous Kinetics similar to K-Blue Aqueous 36 months when stored at 2-8°C
  • Clearly indicates when substrate and acid stop have been added to the wells (turns pink)
  • Ideal for sandwich-based HRP assays
  • 100% solvent free
  • RTU (ready-to-use)
  • Low background
  • High consistency between lots
  • 200 mL
  • 500 mL
  • 1 L
  • 20 L (1 x 20 L)

For research and manufacturing use only.