Quantitative ELISA for Bt-Cry1Ab/1Ac
Introduction:
B. thuringiensis forms crystals of proteinaceous insecticidal δ-endotoxins (Cry toxins) which are encoded by cry genes. Many genetically engineered crops have been developed with insect tolerance till date by expressing cry genes from Bacillus thuringiensis. Bt-modified crops appear to be safe for farmers and consumers.
Intended Use:
Bt-Cry1Ab/1Ac ELISA assay is used as an analytical tool for quantitative laboratory screening of presence or absence of Bt-Cry1Ab/1Ac proteins in leaves, seedlings and seeds of transgenic crop varieties like cotton. The assay can be used to detect the presence of these proteins in cotton products including Bollgard™ and Widestrike™.
Principle:
The Bt-Cry1Ab/1Ac Quantitative kit is an Enzyme-Linked Immunosorbent Assay (ELISA). Plant leaf or seedsample extracts are added to microtiter plate, which is coated with antibodies to Cry1Ab/1Ac toxin. If Bt-Cry1Ab/1Ac protein is present in the sample, it binds to the antibody and thus is captured in the wells, which can then be detected by addition of an Enzyme Conjugate. This immunological reaction results in the formation of a sandwich complex of solid phase Antibody-Toxin-Enzyme labeled antibody. The wells are washed to remove any unbound reactants as per the wash procedure. The Substrate is then added. If any enzyme is present, the substrate will be hydrolysed signifying the presence of Bt-Cry1Ab/1Ac protein. This color reaction can be measured spectrophotometrically or observed visually. The color development is proportional to the concentration of Cry1Ab/1Ac toxin in the sample which can be determined by extrapolating on the standard curve.
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