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High Sensitivity ELISA Technology

Low abundance target analysis made easy

High Sensitivity ELISA Protocol

See available High Sensitivity ELISA Kits

High Sensitivity ELISA Assay Setup

Assay set up is similar to a conventional ELISA, using Biotin-streptavidin-HRP, but includes the addition of an amplification reagent I (contains biotinyl-tyramide).

High Sensitivity ELISA Using Biotin Streptavidin HRP

The innovative key substance here is biotinyl-tyramide that is added to the preparaton after streptavidin biotin peroxidase complex. The trick lies in the enzyme HRP using the biotinyl-tyramide as a substrate and catalyzing it from soluble to a highly reactive intermediate that  binds covalently to any protein in the well, e.g. blocking proteins but predominantly in close proximity to the peroxidase. The Result: Additional biotin- binding sites in the area of the relevant antigen/HRP. The amount of reacted biotinyl-tyramide is proportional to the amount of HRP in the well.

High Sensitivity ELISA Multiplying HRP

After removal of excess biotin-Tyramide from the wells, streptavidin-HRP (Amplification Reagent II) is added, and binds to the additional biotin sites created during the biotin-Tyramide reaction. This amplification step effectively multiplies the quantity of HRP molecules for the final substrate reaction.

High Sensitivity ELISA Key

High Sensitivity ELISAs are based on a patent from PerkinElmer LAS, patent number 5,731,158; 5,583,001; 5,196,306.