Chemiluminescence immune detection reagent kit for detecting ractopamine
A technique of chemiluminescence immunity and ractopamine, applied in the field of immunological detection, can solve problems such as prolonging detection time, achieve the effects of increasing sensitivity, simple instrument, and ensuring long-term effectiveness
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Embodiment 1
[0049] 1) Preparation of each component of the kit
[0050] Preparation of ractopamine hapten: acidify ractopamine, react with sodium nitrite in a low-temperature environment without light at 4°C, and generate intermediates containing diazonium cations. The diazotized ractopamine was used as a hapten for subsequent synthesis of immune antigens and coating antigens.
[0051] Synthesis of bovine serum albumin-ractopamine immune antigen: the immune antigen was obtained by coupling ractopamine and bovine serum albumin (BSA) by diazotization method.
[0052] Synthesis of human serum albumin-ractopamine-coated antigen: the immunization antigen was obtained by coupling ractopamine and human serum albumin (HSA) by a diazotization method.
[0053] Preparation of ractopamine-peroxidase-labeled antibody: inject the above-mentioned immunization antigen into Balb / C mice, and make it produce antibody serum after several booster immunizations. Take out the splenocytes of the immunized mice...
Embodiment 2
[0081] The chemiluminescent immunoassay kit for detecting ractopamine includes the following components:
[0082] (1) 48-well opaque white ELISA plates (6 strips x 8 wells) are coated with ractopamine-mouse serum albumin cross-linked complex, and are vacuum-sealed with aluminum film.
[0083] (2) 5 bottles of ractopamine standard solution, the concentrations are respectively:
[0084] 0μg / L, 0.04μg / L, 0.36μg / L, 3.24μg / L, 29.16μg / L
[0085] (3) Ractopamine-horseradish peroxidase labeled antibody solution.
[0086] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).
[0087] (5) 50 times concentrated buffer. Diluted 50 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L Tris-HCl buffer, pH6.9, containing 0.1% (v / v) Tween-20.
Embodiment 3
[0089] The chemiluminescent immunoassay kit for detecting ractopamine includes the following components:
[0090] (1) 96-well opaque white ELISA plate (12 strips x 8 wells) coated with ractopamine-egg albumin cross-linked complex, and vacuum-sealed with aluminum film.
[0091] (2) 6 bottles of ractopamine standard solution, the concentrations are respectively:
[0092] 0μg / L, 0.05μg / L, 0.25μg / L, 1.25μg / L, 6.25μg / L, 31.25μg / L.
[0093] (3) Ractopamine-horseradish peroxidase labeled antibody solution.
[0094] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).
[0095] (5) 10 times concentrated buffer. Diluted 10 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L glycine-HCl buffer, pH6.9, containing 0.02% (v / v) Tween-20.
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