Indirectly competitive enzyme-linked immunosorbent assay kit for fenvalerate residual
An enzyme-linked immunosorbent adsorption and fenvalerate technology, which is applied to the analysis of materials, biological testing, material inspection products, etc., can solve the problems of complex operation, low sensitivity, and unstable detection results, and achieve less time-consuming, pre-treatment The effect of simple process
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Embodiment 1
[0019] Embodiment 1 Kit operation and result calculation
[0020] Kit operation and result calculation: After the sample to be tested is pretreated, it is made up to volume with PBS containing 50% methanol for later use. Unpack the vacuum packaging bag and take out the microtiter plate, and equilibrate at room temperature for 5 minutes for later use. Prepare 0mg / L, 0.05mg / L, 0.5mg / L, 5mg / L, 50mg / L, 100mg / L, 200mg / L fenvalerate standard solution, add 50μL standard sample or processed sample to each well 2 to 4 repetitions of the standard sample and sample, add 50 μL of diluted antibody, and incubate at 37°C for 30 minutes; pour out the liquid in the well, wash 2 to 6 times with diluted PBST, and invert the microtiter plate on a water-absorbing Pat dry on the paper; add 100 μL enzyme-labeled goat anti-rabbit secondary antibody diluted at 1:1000, incubate at 37°C for 30 minutes; pour out the liquid in the well, wash the plate 2-6 times with PBST, and pat dry; take liquid A Mix ...
Embodiment 2
[0022] Example 2 Synthesis of 3-[(±)cyano[(CIS)2-[3-chlorophenyl]3-methylbutylcarbonyloxy]phenoxy]phenylpropionic acid
[0023] Synthesis of 1-bromo-3-(dimethoxymethyl)benzene
[0024] Put 18.5g (0.1mol) of 3-bromobenzaldehyde in a 100mL three-necked flask, add 5mL of methanol, stir at room temperature for about 2min, then add dropwise 12.2g (0.118mol) of trimethyl orthoate and 10mL of Mixed solution of methanol, after stirring evenly, add 2 drops of concentrated sulfuric acid dropwise, at this time the system is slightly exothermic. Stir at room temperature, and monitor the progress of the reaction by TLC. After the reaction is complete, dilute the reaction system with 40mL of ether, and then use 10mL of 5% Na 2 CO 3 Wash the organic phase with an aqueous solution. After shaking, a large amount of white solids precipitate out, and the interface between the organic phase and the water phase is unclear. Add 30 mL of deionized water to wash the organic phase, then wash with 1...
Embodiment 3
[0045] Example 3 Preparation of polyclonal antibody to fenvalerate
[0046] Conjugate the hapten with bovine serum albumin by the active ester method, dissolve 2mg of the conjugate in 1mL of normal saline, mix it with 1mL of complete Freund's adjuvant, inject it into the thigh of New Zealand big-eared white rabbit after fully emulsifying, and then inject it into the thigh of New Zealand white rabbit every other day Immunization was boosted once every two weeks, and incomplete Freund's adjuvant was used to mix with the immunogen. The immunization site was subcutaneous on the back of the neck. From the third immunization, blood was collected from the rabbit ear vein one week after each immunization to test the serum titer. A total of 5 times of immunization, the whole blood was collected from the rabbit carotid artery one week after the last immunization, and the rabbit antiserum was crudely extracted by 35% saturated ammonium sulfate salting out method, and finally purified by D...
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