When performing ELISAs, a soluble substrate is used to generate a signal in solution. Colorimetric substrates produce a colored end product and chemiluminescent signals emit light that is measured by a luminometer.
KPL offers a variety of standard and proprietary colorimetric substrates for ELISA applications that can be used with horseradish peroxidase. They are tested to assure lot-to-lot consistency and reproducibility. As stable liquid systems they offer convenience and a range of sensitivity options.
Colorimetric Peroxidase Substrates
SureBlue™ and SureBlue Reserve™ TMB substrates are offered as fast-acting, ready-to-use solutions. Our top tier SureBlue Reserve™ is the substrate of choice when highest sensitivity is required. TMB Peroxidase Substrate is economically priced as a stable two-component solution and offers performance equivalent to SureBlue. All three TMB substrates turn blue upon reaction with peroxidase and are capable of both kinetic and endpoint determinations. ABTS® Substrate is a dependable, tried-and true substrate that is ideal when moderate sensitivity is acceptable and low background is essential. It is supplied as either a one- or two-component formulation that produces a blue-green color.
See graph below for performance comparison.
Chemiluminescent Peroxidase Substrates
Microplate-based chemiluminescence ELISA systems are becoming popular due to their increased sensitivity over conventional colorimetric methods, reaction speed and simplified procedures (no stopping reagent required). KPL offers two HRP chemiluminescent substrates for ELISA, LumiGLO® HRP Substrate and LumiGLO ReserveTM HRP Substrate. Both substrates are mix-and-use substrates with extended linear detection ranges and long glow times. LumiGLO detects peroxidase at the picogram level, and LumiGLO Reserve enables femtogram-level sensitivity.