Time-resolved fluorescence immunoassay kit for detecting malachite green
A time-resolved fluorescence and malachite green technology, applied in biological testing, fluorescence/phosphorescence, analytical materials, etc., can solve the problems of complex sample pretreatment, high detection cost, and expensive instruments, and achieve simple operation, wide application, and specificity strong effect
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[0022] 1. Preparation of immunogen and coating agent
[0023] Synthesis of the immunogen (malachite green-BSA) of the present invention: Accurately weigh 324mg of malachite green and dissolve in 2mL N,N-dimethylformamide, add γ-aminobutyric acid solution drop by drop under stirring, and stir for 3 Hours, adjust the pH of the reaction solution to about 10. The precipitate was removed by centrifugation. Add the above reaction dropwise to the BSA solution (320mg BSA dissolved in 5mL physiological saline), then add 23mg of N-hydroxysuccinimide (NHS), 45.4mg of N,N-dicyclohexylcarbodiimide (DCC), React overnight at 4°C, remove the precipitate by centrifugation, take the supernatant and dialyze it with phosphate buffered solution (PBS) for 3 days, change the dialysate every 6 hours, freeze-dry the obtained product, and store it at -20°C for later use;
[0024] Synthesis of coating source (malachite green-OVA): In the above reaction, after replacing BSA with OVA, the reaction conju...
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