Carbendazim monoclonal antibody, preparation method and application thereof
A technology of monoclonal antibody and carbendazim, applied in biochemical equipment and methods, chemical equipment and methods, methods based on microorganisms, etc., can solve the problems of failing to meet EU standards
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Embodiment 1
[0034] Example 1 Obtaining the Hybridoma Cell Line MC-3 CGMCC No. 4575 and Identification of the Anti-Carbendazim Monoclonal Antibody It Produced
[0035] 1. Obtaining the hybridoma cell line MC-3 CGMCC No. 4575
[0036] 1. Amino modification of carbendazim
[0037] The carbendazim was amino-modified by the ethylene glycol method. The specific method is as follows: Dissolve 5 mg carbendazim in 0.65 mL dimethylformamide, add 350 μL 0.05 mmol / L sodium periodate within 2 minutes, react in the dark for 30 minutes at room temperature, add 130 μL 1mol / L ethylene di Alcohol, shake well for 5 minutes to get amino-modified carbendazim ML.
[0038] 2. Preparation of carbendazim complete antigen ML-KLH
[0039] Take 1130 μL of amino-modified carbendazim ML, add 650 μL of 15 mg / mL keyhole limpet hemocyanin, that is, KLH (preparation buffer is 0.1 mol / L sodium bicarbonate at pH 8.2), react at room temperature for 1 hour, add total 0.7 times the volume of 0.4 mol / L sodium borohydride ...
Embodiment 2
[0073] Example 2 Establishing an indirect competitive ELISA method with the monoclonal antibody secreted by the hybridoma cell line MC-3
[0074] 1. Preparation of carbendazim standard substance: carbendazim standard substance, using high-purity water to prepare standard solutions with the following concentrations (in ppm): 10, 5, 1, 0.5, 0.1, 0.
[0075] 2. Antibody solution preparation: the monoclonal antibody prepared in Example 1 was labeled with horseradish peroxidase, and then the horseradish peroxidase-labeled monoclonal antibody was diluted with phosphate buffer to a working concentration of 1:20000, Then add 1% bovine serum albumin (BSA).
[0076] 3. Preparation of washing solution (10×PBST): 0.1mol / L pH7.5 phosphate buffer containing 0.2% (volume percentage) Tween-20 and 80g / L sodium chloride. When in use, dilute the solution 10 times with pure water before use.
[0077] 4. Substrate solution preparation B solution: use 0.1mol / L sodium acetate-citric acid buffer...
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