TMB Reagent – Assistive Reagent

Store at 2-8°C for 2 years.

TMB Substrate (1-Component) for ELISA

1. Add 100ul of this product into each well.
2. Cover the plate and incubate at 37°C in dark within 30min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated HRP), the other wells show no obvious color.
3. Add 100ul of the stop solution 0.1% NaF or 0.3M H₂SO₄ or 0.25M HCl into each well.
4. Read the O.D Absorbance at 650nm if use 0.1% NaF as the stop solution, or at 450nm if use 0.3M H₂SO₄ or 0.25M HCl as the stop solution.

1. This product is sensitive to oxidizng agent, end users should ensure to avoid any possible contamination.
2. Use this product in dark, it is better to use the plastic containers to subpackage, do NOT use the metal or contaminated containers.
3. High concentration of HRP may lead to the TMB turns to green or red, or produce sediment, in this way, end users should decrease the amount of antibodies or HRP conjugations or reduce the incubation time.