Deoxynivalenol, DON ELISA Kit

Deoxynivalenol

Themethod is based on an indirect competitive ELISA assay. The drug of interesthas been coated in the plate wells. During the analysis, sample is addedalong with the primary antibody specific for the target drug. If the targetis present in the sample, it will compete for the antibody, therebypreventing the antibody from binding to the drug attached to the well. Thesecondary antibody, tagged with a peroxidase enzyme, targets the primaryantibody that is complexed to the drug coated on the plate wells. Theresulting color intensity, after addition of substrate, has an inverserelationship with the target concentration in the sample.

Equipment: 1.Validated microplate reader. 2.Homogenizer 3.Electronic balance 4.Centrifuger 5.Shaker for microtiter plates (optional) 6.Organomation 7.Vortex genie 8.Validated adjustable micropipettes, single and multi-channel. 9.Timer

Homogenized, centrifuged, evaporated, diluted for assay.

Wheat, corn, feed, Beer, and wort

1.Add 50ul of thestandard solutions or samples (sample extracts) into the wells of the teststrips according to the working scheme given. We recommend using duplicatesor triplicates. 2.Add 50ul ofantibody solution to the individual wells successively using a multi-channelpipette or a stepping pipette. Incubate the strips for 30 minutes at roomtemperature. Wash the strips four times using the 1X washing buffer solution.Use at least a volume of 250 ul of washing buffer for each well andeach washing step. Remaining buffer in the wells should be removed by pattingthe plate dry on a stack of paper towels. 3.Add 100ul ofantibody solution to the individual wells successively using a multi-channelpipette or a stepping pipette. Cover the wells with parafilm or tape and mixthe contents by moving the strip holder in a circular motion on the benchtopfor about 30 seconds. Be careful not to spill contents. 4.Incubate the strips for 30 minutes at room temperature. 5.After incubation, remove the covering and vigorously shake the contents ofthese wells into a sink. Wash the strips four times using the 1X washingbuffer solution. Use at least a volume of 250 ul ofwashing buffer for each well and each washing step. Remaining buffer in thewells should be removed by patting the plate dry on a stack of paper towels. 6.Add 100 ul of TMB Solution into each well. Cover the wells with parafilm ortape and mix the contents by moving the strip holder in a circular motion onthe benchtop for about 30 seconds. Incubate the strips for 5-10 minutes atroom temperature. Protect the strips from direct sunlight. 7.Add 50 ul of stopsolution to the wells in the same sequence as for the substrate solution. 8.Read the absorbance at 450 nm using a microplate ELISA photometer within 5minutes after the addition of the stopping solution.