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Indirect-competition-law enzyme linked immunosorbent assay kit for detecting phenylethanolamine A

An enzyme-linked immunosorbent reagent and phenylethanolamine technology, which is applied in measuring devices, instruments, scientific instruments, etc., to achieve the effects of low requirements for instruments and equipment, high accuracy and strong specificity

Inactive Publication Date: 2012-06-27
于洪侠
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ELISA method of phenylethanolamine A has not been reported yet

Method used

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  • Indirect-competition-law enzyme linked immunosorbent assay kit for detecting phenylethanolamine A
  • Indirect-competition-law enzyme linked immunosorbent assay kit for detecting phenylethanolamine A
  • Indirect-competition-law enzyme linked immunosorbent assay kit for detecting phenylethanolamine A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Synthesis of Immunogen and Preparation of Immune Serum

[0023] 1.1 Reagents and instruments

[0024] Phenylethanolamine A (gifted by Hangzhou Dean Technology Co., Ltd.), succinic anhydride (purchased from Beijing Yaobei Biotechnology Co., Ltd.), pyridine (Pyridine, AR.WM=79.10, content > 99.5%, developed by Tianjin Kemiou Chemical Reagent center), N, N-dimethylformamide (Dimethylformamide, DMF, produced by ACROSORGANICS, New Jersey, USA, purchased from Bailingwei Chemical Reagent Company), EDC (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.

[0025] Double-beam ultraviolet-visible spectrophotometer (TU-1909, Beijing Puxi General Instrument Co., Ltd.), chromatography device (3057 portable recorder, Chongqing Chuanyi No. 4 Factory; SBS series numerical control drop counter, constant flow pump, automatic p...

Embodiment 2

[0033] Example 2 Establishment of immunoassay method

[0034] 2.1 ELISA method to determine the optimal coating concentration

[0035] Coat the enzyme plate with 100 μl per well of ovalbumin (OVA)-phenylethanolamine A conjugate at a series concentration of 1000 μg / ml, 100 μg / ml, 10 μg / ml, 5 μg / ml, 1 μg / ml, and 0.25 μg / ml , coated at 4°C for 24h, washed 5 times, patted dry, blocked with 200μl blocking solution per well for 12h at 4°C, washed 3 times, and patted 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 take ...

Embodiment 3

[0049] Example 3 Establishment of an enzyme-linked immunosorbent assay kit for detecting phenylethanolamine A

[0050] An enzyme-linked immunosorbent assay kit for detecting phenylethanolamine A was set up to include the following components:

[0051] (1) a microtiter plate coated with phenylethanolamine A antigen;

[0052] (2) Anti-phenylethanolamine A polyclonal antibody with a protein concentration of 1 mg / L;

[0053] (3) goat anti-rabbit anti-antibody labeled with horseradish peroxidase;

[0054] (4) 6 bottles of phenylethanolamine A standard solution, the concentrations are 0 μg / L, 1 μg / L, 5 μg / L, 10 μg / L, 50 μg / L, 100 μg / L;

[0055] (5) Substrate chromogenic solution A liquid is carbamide peroxide, and substrate chromogenic solution B liquid is tetramethylbenzidine;

[0056] (6) The washing liquid is a phosphate buffer containing 0.05% Tween 20;

[0057] (7) The concentrated sample diluent is a phosphate buffered saline solution of 0.1% Tween-20;

[0058] (8) The st...

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Abstract

The invention discloses an indirect-competition-law enzyme linked immunosorbent assay kit for detecting phenylethanolamine A, which comprises a phenylethanolamine A specific antibody, phenylethanolamine A and conjugates and DON-HRP-Conjugated of carrier protein. The indirect-competition-law enzyme linked immunosorbent assay kit for detecting phenylethanolamine A is sensitive, fast and accurate and is mainly used for screening examination of large quantities of samples. Main reagents in the assay kit are all provided in the form of working solution. The indirect-competition-law enzyme linked immunosorbent assay kit for detecting phenylethanolamine A has the advantages of being convenient to use, high in specificity, sensitivity, accuracy and the like and capable of fast detecting residual phenylethanolamine A in feedstuff and animal products.

Description

technical field [0001] The invention relates to the fields of ELISA and veterinary drug residue detection. Specifically, the present invention relates to an enzyme-linked immunosorbent assay kit for detecting phenylethanolamine A. technical background [0002] Phenylethanolamine A, also known as clenrapmine CLRA, is a kind of artificial Synthetic chemicals. Clonpamine, an isomer of formoterol, is Eli Lilly synthesis Ractopamine The by-product of Clenbuterol has the same action and effect as Clenbuterol and Ractopamine. It is a kind of β-agonist and has the effect of nutrient redistribution. It was listed as a substance prohibited from being used in feed and animal drinking water by the Ministry of Agriculture Announcement No. 1519. Phenylethanolamine A and "Clenbuterol" belong to β-adrenoceptor agonists. At the end of 2010, the Ministry of Agriculture announced that it was prohibited Add the substance to drinking water. [0003] Most of the existing detection methods ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 于洪侠
Owner 于洪侠
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