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AFLATOXIN M1
MICROWELL ELISA FOR MILK AND CHEESE
The HELICA Aflatoxin M1 Assay 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
interpretative result is determined.
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