Unlock instant, AI-driven research and patent intelligence for your innovation.

Enzyme-linked immunoassay detection reagent kit for human insulin-like growth factor II

A technology of enzyme-linked immunoassay and detection kit, which is applied in the direction of biological testing, material inspection products, etc., can solve the problems of high price, pollution and radiation, and achieve the effect of sensitive detection

Inactive Publication Date: 2014-03-05
GENERAL HOSPITAL OF PLA
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Insulin-like growth factor Ⅱ radioimmunoassay kit used by Beijing Northern Biological Products Research Institute, but because the marker is radioactive 125I, there are shortcomings of pollution and radiation in use; there are ELISA kits for insulin-like growth factor Ⅱ in foreign countries, but expensive

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Enzyme-linked immunoassay detection reagent kit for human insulin-like growth factor II
  • Enzyme-linked immunoassay detection reagent kit for human insulin-like growth factor II
  • Enzyme-linked immunoassay detection reagent kit for human insulin-like growth factor II

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Preparation of Human Insulin-like Growth Factor II ELISA Detection Kit

[0016] 1. Materials

[0017] Three big-eared white rabbits for immunization, male, weighing 1.5 kg, were provided by the Animal Experiment Center of 301 Hospital. Human IGF Ⅱ polypeptide was purchased from Beijing Boaosen Biotechnology Co., Ltd., and human insulin-like growth factor Ⅱ standard product was purchased from sigma company.

[0018] 2. Preparation and purification of human insulin-like growth factor II antibody

[0019] IGFⅡ and thyroglobulin (TG) were fed according to the optimal molecular ratio (1:100), connected by glutaraldehyde method, subpackaged after dialysis, and freeze-dried to prepare IGFⅡ-TG.

[0020] For the first immunization, IGFⅡ-TG 200 μg and Freund’s complete adjuvant 4ml were fully mixed, and injected into the back of 3 rabbits at multiple points, and boosted immunization once every 4 weeks (100 μg of immunogen and Freund’s incomplete adjuvant 4ml Mix well...

Embodiment 2

[0028] Embodiment 2 detection method

[0029] 1. Preparation of solid-phase antibody microtiter plate

[0030] Take the purified IGFⅡ antibody, prepare 10ug / ml with 0.05M citrate buffer, add 200μl per well into 96-well ELISA plate, incubate at 37°C for 3h, wash the plate, add 3% bovine serum Put albumin in Tris.HCl blocking solution overnight at 4°C, shake off the blocking solution, use a freeze dryer to remove excess water in the well, put it into a tin foil bag and vacuum seal it with a vacuum packaging machine, and store at 4°C.

[0031] 2. Preparation of reagents and instruments

[0032] 1 bottle of freeze-dried human insulin-like growth factor Ⅱ standard substance: each bottle is dissolved in 0.5mL of experimental buffer and the concentration is 250, 83, 27, 9, 3, 1 ng / mL; biotinylated human insulin-like growth factor Ⅱ 1 bottle; 1 bottle of avidin-labeled horseradish peroxidase working solution; chromogenic reagent A: 1 bottle; chromogenic reagent B: 1 bottle, chromoge...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an enzyme-linked immunoassay detection reagent kit for human insulin-like growth factor II and belongs to the technical field of enzyme-linked immune detection. The reagent kit consists of a solid-phase ELISA (Enzyme-linked Immunosorbent Assay) plate, a human insulin-like growth factor II antibody, a human insulin-like growth factor II standard substance, a biotinylated human insulin-like growth factor II, an avidin-marked HRP (Horse Radish Peroxidase) enzyme and an auxiliary reagent. The detection reagent kit can sensitively detect the difference of IGF (Insulin-like Growth Factor) II in serum.

Description

technical field [0001] The invention belongs to the technical field of ELISA detection, in particular to an ELISA detection kit for human insulin-like growth factor II. Background technique [0002] Human insulin-like growth factor Ⅱ (Insulin-Like Growth Factor Ⅱ, IGF Ⅱ) is a single-chain polypeptide with 67 amino acids, with a molecular weight of 7Kd and a structure similar to insulin. Most of it is synthesized by the liver and mainly exists in the blood. The specific binding of receptors can promote cell reproduction, differentiation, growth, metabolism and other biological effects. In recent years, a large number of studies have shown that insulin-like growth factor Ⅱ can promote cell proliferation and malignant transformation. Dysregulation of the factor II gene. At present, there is no report on the research and use of human insulin-like growth factor Ⅱ enzyme immunoassay-related methodological detection and kits in China. Insulin-like growth factor Ⅱ radioimmunoassa...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
Inventor 颜光涛薛辉
Owner GENERAL HOSPITAL OF PLA