Preparing method of test kit special for enzyme linked immunosorbent assay adsorption of vomitus toxin
An enzyme-linked immunosorbent (ELISA) and vomitoxin technology, applied in the biological field, can solve the problems of requiring specialized technicians, expensive detection costs, unfavorable promotion and application, etc., and achieve the effects of simple operation, rapid detection, and convenient use.
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Embodiment 1
[0020] The coating plate is coated with vomitin solid-phase antigen, which uses 96 or 48 or 24-well microtiter plates, with 10mmol / L pH9 Na 2 CO3-NaHCO 3 As the coating solution, dilute DON-OVA to 1.0μg / mL, add 100μl to each well of a 96 or 48 or 24-well microtiter plate, place overnight at 8°C, discard the coating Wash 4, add 5% bovine serum albumin, pH 7.5 acid buffer solution, block overnight at 2°C, discard the blocking solution, blow dry, seal the slats and store at -20°C for storage;
[0021] Said DON standard product is obtained by diluting from pure DON, the diluent is deionized water, and the concentration of DON is 0.001ng / mL;
[0022] Preparation of vomitin-bovine serum albumin (DON-BSA) conjugate: Dissolve DON in pyridine, add 1-butylboric acid to the reaction flask; stir the mixture overnight at room temperature to produce 7,15-oxygen-one (Butylboron)-deoxynivalenol; add succinic anhydride to it, and pass nitrogen into it; seal the reaction flask, and stir the mixture ...
Embodiment 2
[0027] The coating plate is coated with the vomitin solid phase antigen, which uses a 96-, 48-, or 24-well microtiter plate with 100mmol / L pH10Na 2 CO 3 -NaHCO 3 As the coating solution, dilute DON-OVA to 0.1μg / mL, add 100μl to each well of a 96 or 48 or 24-well microtiter plate, place overnight at 5°C, discard the coating Wash 2 times, add 1% bovine serum albumin, pH7 phosphate buffer, block overnight at 8°C, discard the blocking solution, blow dry, seal the slats and store at -40°C for freezer;
[0028] Said DON standard product is obtained by diluting from pure DON, the diluent is deionized water, and the concentration of DON is 1000 ng / mL;
[0029] Preparation of vomitin-bovine serum albumin (DON-BSA) conjugate: Dissolve DON in pyridine, add 1-butylboric acid to the reaction flask; stir the mixture overnight at room temperature to produce 7,15-oxygen-one (Butylboron)-deoxynivalenol; then add succinic anhydride, and pass nitrogen into it; seal the reaction flask, and stir the mi...
Embodiment 3
[0034] Said coating plate is coated with vomitin solid phase antigen, which adopts 96 or 48 or 24-well microtiter plate, with 55mmol / L pH9.5 Na 2 CO 3 -NaHCO 3 As the coating solution, dilute DON-OVA to 4.95μg / mL, add 100μl to each well of a 96 or 48 or 24-well microtiter plate, place overnight at 2°C, discard the coating Wash 5 times, add 3% bovine serum albumin, pH8 phosphate buffer, block overnight at 5°C, discard the blocking solution, blow dry, seal the slats and store at -30°C for freezing;
[0035] The DON standard product is obtained by diluting from the pure product of DON, the diluent is deionized water, and the concentration of DON is 500ng / mL;
[0036] Preparation of vomitin-bovine serum albumin (DON-BSA) conjugate: Dissolve DON in pyridine, add 1-butylboric acid to the reaction flask; stir the mixture overnight at room temperature to produce 7,15-oxygen-one (Butylboron)-deoxynivalenol; add succinic anhydride to it, and pass nitrogen into it; seal the reaction flask, an...
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