Skip to main content Skip to navigation
TMB Substrate Reagent Set

BD OptEIA™ TMB Substrate Reagent Set

Product Details
Down Arrow Up Arrow

ELISA (Tested During Development)


The TMB substrate reagent set is designed for use in BD OptEIA™ ELISA sets. Substrate Reagent A, which is clear and colorless, contains hydrogen peroxide. Substrate Reagent B is a colorless to a very light amber solution containing 3,3', 5,5' tetramethylbenzidine (TMB) in an organic solvent.

Preparation And Storage

Store unopened reagents at 2-8°C. Do not use after the expiration date. Discard any remaining working solutions after use. Reagents should be brought to room temperature (20-25°C) prior to use. Allow at least 30 minutes for this process. Avoid prolonged exposure to light, contact with metal, air or extreme temperatures as color may develop.

Recommended Assay Procedures

Prepare a working concentration of TMB substrate solution within 15 minutes prior to use by mixing equal volumes of Substrate Reagent A and Substrate Reagent B (e.g. for one 96-well plate, a 12 mL TMB substrate working solution can be prepared by mixing 6 mL of Substrate A with 6 mL of Substrate Reagent B).  When mixed together, TMB substrate solution should be colorless to a very faint blue. In the presence of peroxide-labeled conjugates, the solution should develop a definitive blue color. After the addition of sulfuric or phosphoric acid to stop the reaction, the color changes from blue to yellow.

Danger:  Substrate Reagent B (component 51-2607KC) contains 33.05% methanol (w/w), 13.34% glycerin (w/w), and 1.745% DMSO (w/w).

Hazard statements

Flammable liquid and vapour.

Toxic if swallowed, in contact with skin or if inhaled.

Causes damage to the central nervous system. Route of exposure: Oral.

Causes damage to organs.

Precautionary statements

Keep away from heat/sparks/open flames/hot surfaces. - No smoking.

Keep container tightly closed. Ground and bond container and receiving equipment. Use explosion-proof [electrical/ventilating/lighting/] equipment. Use non-sparking tools. Take action to prevent static discharges. Use suitable extinguishing media to extinguish.

Wear protective gloves / eye protection.

Wear protective clothing.

Do not breathe mist/vapours/spray.

Use only outdoors or in a well-ventilated area.

Wash thoroughly after handling. Do not eat, drink or smoke when using this product.

IF ON SKIN (or hair): Remove/Take off immediately all contaminated clothing. Rinse skin with water/shower.Take off immediately all contaminated clothing. Wash contaminated clothing before reuse.

IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable for breathing.

IF SWALLOWED: Call a POISON CENTER/doctor/ if you feel unwell. Rinse mouth.

IF exposed or concerned: Call a POISON CENTER/doctor/

Store in a well-ventilated place. Keep cool.  Store locked up.  Keep container tightly closed.  

Dispose of contents/container to an appropriate treatment and disposal facility in accordance with applicable laws and regulations, and product characteristics at time of disposal.

Product Notices

  1. Please refer to for technical protocols.
  2. Please refer to to access safety data sheets (SDS).
555214 Rev. 6
Down Arrow Up Arrow
Description Quantity/Size Part Number EntrezGene ID
Substrate Reagent A 300 mL (1 ea) 51-2606KC N/A
Substrate Reagent B 300 mL (1 ea) 51-2607KC N/A
555214 Rev. 6

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.