One-Component TMB ELISA Substrate

TMB is the most popular chromogenic substrate for HRP detection in ELISA.

The One-Component TMB ELISA Substrate is a Ready-to-Use substrate that requires no preparation before use. HRP can catalyze TMB substrate to yield a blue color, the maximal absorbance is at 370 nm or 620-652 nm; however, upon addition of the stop solution, the solution turns to yellow and can be measured at 450 nm.         

Features

Instructions For Use

1. Run ELISA according to the specific protocol through the conjugate incubation step.

2. Wash the wells 3 times with PBST (PBS containing 0.05% Tween-20), 5 minutes each time.

3. Add 100 µL of TMB Substrate (prewarmed to room temperature) to each well. Incubate for 10-60 minutes. Monitor the color intensity.

4. Add 50 µL of stop solution (1M H₂SO₄) to each well.

5. Measure the absorbance at 450 nm within 1 hour.

Application

Peroxidase-based enzyme immunoassays (ELISA)