TMB substrate is used in ELISA procedures, which utilize horseradish peroxidase conjugates. TMB substrate develops a soluble blue reaction product that can be read at 370 or 655 nm at 370 or 655 nm. The reaction is stopped with acid, forming a yellow reaction product which enables accurate intensity measurement at 450 nm.

This kit contain:
Solution A (1% TMB in DMSO) 250mL
Solution B (33% H2O2) 250mL

To prepare a working solution for ELISA, dilute 1.0 mL of Solution A with 9.0 mL of phosphate-citrate buffer pH 5.2, and add 2.0 μL of Solution B. This solution is now ready to be used for ELISA involving an HRP-conjugated Ab.