ELISA reagent kit for detecting melamine
An enzyme-linked immunosorbent reagent and melamine technology are applied in measuring devices, instruments, and material analysis through optical means to achieve the effects of time saving, high accuracy, and no radioactive isotope pollution
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0024] The synthesis of embodiment 1 immunogen and the preparation of immune serum
[0025] 1.1 Reagents and instruments
[0026] Melamine (Beijing Bailingwei Reagent Co., Ltd.), succinic anhydride (purchased from Beijing Yaobei Biotechnology Co., Ltd.), pyridine (Pyridine, AR.WM=79.10, content > 99.5%, Tianjin Kemiou Chemical Reagent Development Center), N , N-dimethylformamide (Dimethylformamide, DMF, produced by ACROSORGANICS, New Jersey, U.S., purchased from Bailingwei Chemical Reagent Company), tributylamine (Tributylamine, produced by ACROSORGANICS, New Jersey, U.S., purchased from Bailingwei Chemical Reagent Company), chlorine Isobutyl formate was purchased (from Bailingwei Chemical Reagent Company), bovine serum albumin (BSA), ovalbumin (OVA), etc. (purchased from Beijing Yaobei Biotechnology Co., Ltd.), and other reagents were of analytical grade.
[0027] Double-beam ultraviolet-visible spectrophotometer (TU-1909, Beijing Puxi General Instrument Co., Ltd.), chromato...
Embodiment 2
[0035] Embodiment 2 The establishment of immunoassay method
[0036] 2.1 ELISA method to determine the optimal coating concentration
[0037] Ovalbumin (OVA)-melamine conjugates with serial concentrations of 1000 μg / ml, 100 μg / ml, 10 μg / ml, 5 μg / ml, 1 μg / ml, and 0.25 μg / ml were used to coat 100 μl per well of the microtiter plate, 4 Coat for 24 hours at ℃, wash 5 times, pat dry, block with 200 μl blocking solution per well for 12 hours at 4 ℃, wash 3 times, and pat dry. Add 100 μl of 1:500 diluted antiserum, react at room temperature for 2 hours, wash three times, immediately add 100 μl enzyme-labeled goat anti-rabbit antibody, react at room temperature for 30 minutes, wash three times, add 100 μl substrate solution, react at room temperature in the dark for 15 minutes, stop with 50 μl stop solution Reaction, microplate reader detects A value (450nm). At the same time, set blank control wells (without adding antiserum, only adding its diluent) and parallel repeat wells, and ...
Embodiment 3
[0051] Embodiment 3 detects the formation of the ELISA kit of melamine
[0052] 3.1 Set up an enzyme-linked immunosorbent assay kit for detecting melamine to include the following components:
[0053] (1) Enzyme plate coated with melamine antigen;
[0054] (2) Anti-melamine polyclonal antibody with a protein concentration of 1 mg / L;
[0055] (3) goat anti-rabbit anti-antibody labeled with horseradish peroxidase;
[0056] (4) 6 bottles of melamine standard solution, the concentrations are 0 μg / L, 1 μg / L, 5 μg / L, 10 μg / L, 50 μg / L, 100 μg / L;
[0057] (5) Substrate chromogenic solution A liquid is carbamide peroxide, and substrate chromogenic solution B liquid is tetramethylbenzidine;
[0058] (6) The washing liquid is a phosphate buffer containing 0.05% Tween 20;
[0059] (7) The concentrated sample diluent is a phosphate buffered saline solution of 0.1% Tween-20;
[0060] (8) The stop solution is 2mol / L hydrochloric acid solution.
[0061] 3.2 Set up the enzyme-linked immu...
PUM
| Property | Measurement | Unit |
|---|---|---|
| concentration | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More