TMB is a popular chromogenic substrate for HRP detection in ELISA and is available in several formats. The soluble One-component TMB Substrate are supplied as Ready-to-Use mono-component liquids that require no preparation before use. The product mainly contains 3,3’,5,5’-tetramethylbenzidine and contains no DMF or DMSO.
One-component TMB Substrate yields a blue color (Amax= 370 nm and 652 nm) when reacted with peroxidase in microwell applications. The product changes to yellow (Amax = 450 nm) upon addition of a sulfuric or phosphoric acid stop solution.
Peroxidase-based enzyme immunoassays ELISA is recommended.
This product is stable for 2 years stored at 2°C~8°C. Shading light.
> Economical: Cost less compared with similar products.
> Convenience: mono-component, ready-to-use.
> High sensitivity: Save the dosage of antibody.
> Strong stability: Valid for 2 years at 2~8℃.
> Stable signal: The signal is continuous steady after adding stop solution.
> Safe: Non carcinogenic.
1. Add 100 μL of the TMB Substrate Solution to each microplate well.
2. Incubate for 5~30 min according to the reaction system.
3. Stop reaction by adding 50 uL stop solution (1M HCl or 1M H2SO4).
4. Measure the absorbance of each well at 450 nm.
1. This substrate is light sensitive, avoid direct sunlight during operation.
2. This substrate is sensitive to contamination of oxidizing agent. Avoid oxidizing agent contamination during operation.
3. Never pipette directly from the bottle. Pour out required amount into a tube and pipette from the tube.
4. Do not return excess TMB to primary storage container.