Aflatoxin B1 ELISA Kit 96 Well Plate

The ELISA Aflatoxin B1 Kit allows the quantitative determination of Aflatoxins in food and feed crop (cereals, nuts, etc.). For further applications the user is encouraged to verify the validity of the tests themselves or contact the manufacturer.
The test is easy to use, requires no complex instrumental equipment, and delivers the sensitive results quickly. After a sample preparation to isolate the Aflatoxins, a maximum of 42 samples in duplicate can be tested in 45 minutes with the ELISA Aflatoxin B1 Kit. The sample extraction with methanol/water is described in the following. Alternatively, Aflatoxins can also be isolated with the help of immunoaffinity chromatography.
The assay is carried out in a microplate, of which the cavities have already been pre-coated with a special anti-Aflatoxin antibody.
To carry out the assay, Aflatoxin B1 (AFB1) standards and sample extracts respectively and Aflatoxin B1 peroxidase conjugate (AFB1-HRP-conjugate) are pipetted into the cavities. Aflatoxin from the standard or sample now competes with the HRP labelled Aflatoxin for the antibody binding sites. After an incubation period of 30 minutes, unbound components are removed by rinsing them off and then peroxidase substrate solution (3,3’,5,5’-Tetramethylbenzidine, TMB) is pipetted into all cavities. The AFB1-HRP conjugate that is bound to the antibody reacts with the substrate solution by forming a blue colour. After 15 minutes, this reaction is terminated by adding the stop solution. The colour then changes from blue to yellow, which is detected photometrically. With the help of a microplate reader, the yellow colouring is measured as an optical density (OD) at a wavelength of 450nm (reference value 620nm).
The Aflatoxin concentration is inversely proportional to the colour intensity. The higher the OD measurement, the lower is the concentration of Aflatoxin in the standard or sample.
Assay time 45 min (30+15 min)
Lower detection limit matrices: ≤2 ppb
Cross reactivity: Aflatoxin B1: 100 %; Aflatoxin B2: 63 %; Aflatoxin G1: 65 %; Aflatoxin G2: 19 %; Aflatoxin M1: 7 %;
Storage: 2-8 °C

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