USE PRINCIPLES
The Capsaicin Plate Kit uses a polyclonal antibody that binds both capsaicin and a capsaicin-enzyme conjugate. Capsaicin in the sample competes with the capsaicin-enzyme conjugate for a limited number of antibody binding sites. Antibodies, which bind capsaicin, are immobilized to the inside of the test wells.

In the assay procedure you will:
•   Add a mixture of a sample containing capsaicin and capsaicin-enzyme conjugate to a test well. The conjugate competes with any capsaicin in the sample for the same antibody binding sites.
•   Wash away any unbound molecules, after you incubate this mixture for 10 minutes.
•   Add clear substrate solution to each well. In the presence of bound capsaicin-enzyme conjugate, the substrate is converted to a blue compound. One enzyme molecule can convert many substrate molecules.

Since the same number of antibody binding sites are available in every well, and each well receives the same number of capsaicin-enzyme conjugate molecules, a sample containing a low concentration of capsaicin allows the antibody to bind many capsaicin-enzyme conjugate molecules. The result is a dark blue solution. Conversely, a high concentration of capsaicin allows
fewer capsaicin-enzyme conjugate molecules to be bound by the antibodies, resulting in a lighter blue solution.

NOTE: Color is inversely proportional to capsaicin concentration.
Darker color = Lower concentration
Lighter color = Higher concentration