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Radioimmunoassay kit for measuring rat serum total IgE, and detection method thereof

A radioimmunoassay, analytical reagent technology, applied in measurement devices, analytical materials, instruments, etc., can solve problems such as good measurement repeatability

Inactive Publication Date: 2011-11-16
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The radioimmunoassay for detecting IgE, although the problem of radioactivity hinders its development and wide application, but its measurement repeatability is good, and it is still used in research institutions

Method used

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  • Radioimmunoassay kit for measuring rat serum total IgE, and detection method thereof
  • Radioimmunoassay kit for measuring rat serum total IgE, and detection method thereof
  • Radioimmunoassay kit for measuring rat serum total IgE, and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026]Embodiment 1: the preparation of antibody and separating agent

[0027] 1. Preparation of rabbit anti-rat IgE antibody

[0028] Intralymphatic injection of a small amount of antigen was used for immunization: Freund's complete adjuvant and antigen (rat IgE solution) were mixed and emulsified in a volume ratio of 1:1, and 0.2ml of the mixed emulsion was firstly injected into the front soles of both feet of New Zealand male rabbits. Ten days later, popliteal lymph nodes were seen to be enlarged, and the emulsion mixed with Freund's complete adjuvant and antigen was injected into the enlarged lymph nodes. Thereafter, the antigen was emulsified with Freund's incomplete adjuvant every 14 days, and strengthened twice in the same way. During immunization, the titer of antiserum was monitored by agar immuno-double diffusion method. Serum was taken from the heart two months later, and the polyclonal antibody (IgG) of rabbit anti-rat IgE was obtained by stepwise precipitation wi...

Embodiment 2

[0034] Example 2: Rat IgE 125 I mark

[0035] Take a small reaction test tube coated with lodogen (5 μg, Sigma), add 5 μg / 50 μl rat IgE solution, and 2 μl Na 125 I solution (~500 μCi). After stirring the reaction at room temperature for 5 min, the reaction was terminated with 0.01 mol / L PBS buffer solution of pH 7.4. Then it was purified by Sephadex G25-80 gel dextran chromatography column (1 cm×25 cm), the eluent was 0.01 mol / L PBS buffer solution with pH 7.4, and the flow rate was 1 ml / min. Collect the eluate (1ml / tube) and measure the radioactivity in each tube in turn, and combine the collected 125 I-labeled rat IgE peak,

Embodiment 3

[0036] Embodiment 3: kit composition

[0037] 1. Rat IgE standard solution: Take rat IgE standard products and make standard series with concentrations of 0.5, 2, 6, 18, 50, 180, 500, 1,500 ng / ml, 8 bottles in total, 0.5ml / bottle . Determination of standard point concentration: the correlation coefficient of the fitted standard curve must be greater than 0.99, otherwise the standard concentration should be adjusted.

[0038] 2. Marker solution: 125 I-labeled rat IgE, one bottle, 10ml, red, containing 15,000-30,000 cpm radioactivity per 100ul.

[0039] 3. Antiserum solution: rabbit anti-rat IgE antiserum, one bottle, 10ml, blue. Determination of antiserum dilution: antiserum and 125 The maximum binding rate of I-labeled rat IgE = 35-50% (B 0 value) is the practical dilution.

[0040] 4. Quality control samples: prepared from known rat serum samples, three bottles, 0.5ml / bottle, one bottle each with low, medium and high concentrations, the concentrations are 10, 100 and 50...

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Abstract

The invention discloses a magnetic separating radioimmunoassay kit for measuring rat serum total Ig. The radioimmunoassay kit comprises IgG magnetic nano particles and corresponding reagent. The analysis kit is characterized by having measurement sensitivity reaching 0.5ng / ml, good precision and accuracy, and can be used in measurement of microscale IgE in rat serum. The invention also discloses a method utilizing the radioimmunoassay kit for detection rat serum total IgE.

Description

technical field [0001] The invention relates to a radioimmunoassay kit for detecting total IgE content in rat serum, in particular to a method for preparing magnetic nanoparticles based on coupling goat anti-rabbit IgG; the invention also relates to a radioimmunoassay kit for total IgE content in rat serum Detection method. Background technique [0002] Allergy is one of the most common diseases in modern society, it is caused by different kinds of allergens. About 5-10% of our country, about 5-25% of people in the United States and Europe have suffered from allergic diseases, among which the symptoms of children and adolescents are particularly obvious, and the threat is more serious. Allergic reaction is also called allergic reaction, and allergen is also called allergen. According to the opinion published by the European Academy of Allergy and Clinical Immunology (EAACI), there are two types of immune-mediated allergic reactions: IgE-mediated allergic reactions and non-...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 张彤钟高仁林敏金若敏李秀芳韩向东徐培康
Owner SHANGHAI UNIV OF T C M
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