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User Instruction Store reagents A and B in the darkness at 4-8°C.Mix equal volumes of reagent A and B (chemiluminescent substrate) in a clean container and equilibrate to room temperature 30 minutes before use.Do not contaminate the HRP enzyme substrate with HRP or other proteins. Never pipette directly from the bottle stock or pour aliquoted solution back into the stock bottle.100 l of Agrisera ELISA-ECL should be used/well.Incubate at RT for optimial detection e.g. for example 20 min. after substrate contact. Assay in appropriate microtieter plates designed for chemiluminescence. Detection conditions need to be optimized for each assay system. This reagent is extremely sensitive and very high HRP concentration can "burn out" the substtrate. This substrate has a very wide detection range for HRP enzyme in a soluble environment. HS code for this product is: 3822.00.0002.