TMB substrate(3,3'.5,5'-tetramethylbenzidine) is a chromogen that yields a deep blue color (maximum absorbance at 605nm or 650nm) when oxidized with hydrogen peroxide(catalyzed by HRP). The color then changes to yellow with the addition of 2N H2SO4 with maximum absorbance at 450 nm. Our TMB Substrates(PeroxichromTM, Peroxichrom ExcelTM) are one-component substrates that require no preparation before using, stable and sensitivity. Also PeroxichromTM and Peroxichrom ExcelTM contain no organic solvents such as DMF, DMSO, and methanol so there is no issue of safety with user.
TMB substrate is sensitive to contamination from a variety of oxidizing agents. Avoid prolonged exposure to light, contact with metal or air. We recommend using TMB substrate by pouring out required amount into a reservoir and do not return excess TMB to provided bottle.
Stable at RT for 1 month (Stable at 4℃ for 1 year)
1. Warm to room temperature prior to use if you store TMB solution at 4℃
2. Dispense 1-Component appropriate TMB solution or 100ul into each well
3. After sufficient color development (5-10minutes at room temperature or at 37℃)
Add 50ul Stop Solution (2N H2SO4 or 1M H3PO4) to each well.
4. Read plates at 450nm