Rapid Aflatoxin M1 ELISA Kit

The Sigma-Aldrich^® Aflatoxin M₁ ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid phase competitive enzyme immunoassay. An antibody with a high affinity for aflatoxin M₁ is coated onto polystyrene microwells. Standard or sample is added to the appropriate well and if Aflatoxin M₁ is present it will bind to the coated antibody. Subsequently, aflatoxin bound to horse - radish peroxidase (HRP) is added and binds to the antibody not already occupied by aflatoxin M₁ present in the sample or standard. After this incubation period, the contents of the wells are decanted, washed and an HRP substrate is added which develops a blue color in the presence of enzyme. The intensity of the color is directly proportional to the amount of bound conjugate and inversely proportional to the amount of aflatoxin M₁ in the standard or sample. Therefore, as the concentration of aflatoxin M₁ in the sample or standard increases, the intensity of the blue color will decrease. An acidic stop solution is added which changes the chromogen color from blue to yellow. The microwells are measured optically by a microplate reader with an absorbance filter of 450nm (OD₄₅₀). The optical densities of the samples are compared to the OD′s of the kit standards and an interpolated result is determined.

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