Carbendazim detection ELISA (enzyme linked immunosorbent assay) kit and application thereof
An enzyme-linked immunosorbent reagent, carbendazim technology, applied in the field of enzyme-linked immunosorbent immunoassay kits for the detection of carbendazim, to achieve the effects of efficient detection methods, low detection limits, and convenient portability
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Embodiment 1
[0028] Example 1 The preparation of the component of the box component
[0029] 1. Preparation of polymorrhea semi -antigen
[0030] The polymorrhea ingredients react through the nitrification and introduced nitro on the benzene ring.
[0031] Toto fluoride 20ml plus triathide anhydride 2ml, ice water bath, drop to 0 ° C, add 0.5g of ammonium nitrate, stir 1h for 1h, add a three -fluoroceticolate solution containing a multi -bacterial spirit 1.0g, continue to stir, and react for 2h.Stop the reaction, the neutral and neutral of the dilute sodium hydroxide solution, the dichloromethane extraction, the water washing, the steaming dry, and the crystal of the ether, the compound A0.76g, the income of 67%.1HNMR (CDCL3,300MHz) Δ: 8.31 (1H, DD, J = 1.616, J = 1.239), 7.69 (1H, DD, J = 8.716, J = 1.616), 7.64 (1H, DD, J = 8.716, J =1.239), 3.85 (3h, s).
[0032] The compound A0.7g plus ethanol is dissolved, plus 0.43g of chloride tin solution of 10ml, passes through nitrogen, and adds a re...
Embodiment 2
[0056] Example 2 detect the formation of polymorrhea enzyme linked immune kit
[0057] Establish an enzyme -linked immune kit to detect polymorphic spirit, so that it includes the following components:
[0058] (1) Enzymes of the packets of the bacterial puppet couplet antigen;
[0059] (2) 6 bottles of multi -bacterial spiritual standard products, with the concentration of 0 μg / L, 0.1 μg / L, 0.3 μg / L, 0.9 μg / L, 2.7 μg / L, 8.1 μg / L;
[0060] (3) Polymogenic antibody with spicy root peroxidase marks;
[0061] (4) The substrate color rendering liquid consists of A liquid and liquid, and A solution is peroxidine, and the B liquid is a tetrahydramine aniline;
[0062] (5) The termination solution is 2 mol / L sulfuric acid;
[0063] (6) The washing liquid was pH value of 7.4, and containing 0.5%to 1.0%of the vomiting temperature -20, 0.01 ‰ ~ 0.03 ‰ sodium nitride nitride preservatives, 0.1 ~ 0.3 mol / L phosphate buffer liquid;
[0064] (7) Phosphate buffer with a pH value of 7.0 and 0.02m...
Embodiment 3
[0065] Example 3 Detection of Multiococcus Spirit in Tobacco Leaf
[0066] 1. Pre -sample processing
[0067] Called 1.0 ± 0.05g tobacco leaves to 50ml of polystyrene centhal, add 10ml tobacco leaf extract, and use a uniform pulp to smash it; filter the crushed samples with the filter membrane;Remove ion water and mix it in full; then take the above liquid 50ml and add 950ml to re -soluble work solution to fully mix; take 50ml for analysis.
[0068] 2. Use the kit to detect
[0069] Add the standard solution / sample to 50ml to the corresponding micropore, add 50ml / hole of the antibody working liquid, gently oscillate and mix it, and use the cover plate cover to react to the light environment at a rear of 25 ° C to react in the light.Carefully unveil the cover of the plate, dry the liquid in the hole, use 250ml / hole with the washing work liquid, and wash it 4-5 times in full, each time the interval is 10s, and use the water absorption paper to dry it (the bubbles that are not cleare...
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