UltraScence TMB ELISA Substrate

UltraScence TMB ELISA Substrate
UltraScence TMB ELISA Substrate
UltraScence TMB ELISA Substrate

Catalog numberTM001-B100ML

Size100 ml (50 ml×2)


StorageStable for up to 12 months at 4 °C


TMB ELISA Substrate, known as 3,3',5,5'-tetramethylbenzidine, is a chromagen that detects horseradish peroxidase (HRP) activity when oxidized with hydrogen peroxide and yields a blue color with major absorbances at 370 and 652 nm. The substrate is supplied as a ready to use, mildly acidic solution. The unreacted substrate should be colorless or very light yellow in appearance. The reaction can be stopped using appropriate stop solution, either the sulfuric or phosphoric acid, and producing a soluble yellow reaction product with the absorbance at 450 nm.

UltraScence TMB ELISA Substrate


Unlike other commercially available substrates, Alfa Chemistry's TMB contains no DMF.

UltraScence TMB ELISA Substrate

  • Ready-to-use solutions
  • No hydrogen peroxide required
  • No filtering required
  • Noncarcinogenic
  • Significantly more sensitive than leading manufacturers on the market
Reaction Setup

1. Equilibrate the TMB ELISA Substrate to room temperature before using.
2. Mix of equal volume TMB ELISA Substrate A and B solution and then add 100 μL mixture to each microplate well.
3. Incubate in dark for 20-30 minutes or until desired color develops.
4. Add 100 μL stop solution (0.2~2M sulfuric acid) to each well.
5. Read the absorbance of each well at 450 nm within 15 minutes.


UltraScence TMB ELISA Substrate

Required Materials but Not Provided
  • ELISA plate(s).
  • Stop solution (sulfuric acid, 0.2~2M H2SO4).
  • Absorbance microplate reader.
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