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Adamantine chemiluminiscence kit for detecting mechano-growth factor and E peptide thereof and manufacturing method of kit

An adamantane chemistry and growth factor technology, applied in the field of adamantane chemiluminescence kits, can solve problems such as slow speed and inability to quantify, and achieve the effects of avoiding result deviation, fast detection, and improving work efficiency

Inactive Publication Date: 2018-09-04
SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, only Western blot method is used to detect MGF and MGF-Ct24E, and this Western blot detection is only a qualitative analysis, not quantitative, and the speed is relatively slow. At present, there is no rapid and accurate quantitative detection of MGF and MGF. -Ct24E method or kit

Method used

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  • Adamantine chemiluminiscence kit for detecting mechano-growth factor and E peptide thereof and manufacturing method of kit
  • Adamantine chemiluminiscence kit for detecting mechano-growth factor and E peptide thereof and manufacturing method of kit
  • Adamantine chemiluminiscence kit for detecting mechano-growth factor and E peptide thereof and manufacturing method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 The preparation method of the chemiluminescent kit for detecting force growth factor and / or its E peptide

[0025] 1) Preparation of streptavidin-coupled magnetic particle working solution

[0026]The streptavidin-coupled magnetic particle suspension was magnetically separated to remove the supernatant, and the pH was 6.8, and the Tris buffer with a concentration of 0.1 mol / L was resuspended to a concentration of 0.5 mg / mL, and the buffer contained The mass percent content is 0.5% by weight of bovine serum albumin, 0.05% by weight of Triton X-100, 0.05% of proclin 300 and 0.05% of sodium azide.

[0027] 2) Preparation of biotin-labeled MGF antibody

[0028] Take 1mg MGF mouse monoclonal antibody, add appropriate amount of PBS buffer (0.05M, pH7.0-7.6) to adjust the total antibody concentration to 1mg / ml, and put it into a dialysis bag for dialysis, changing the dialysate every 3-4 hours , replace 3-4 times, transfer the antibody to a centrifuge tube or a cry...

Embodiment 2

[0045] Example 2 The preparation method of the chemiluminescence kit for detecting force growth factor and / or its E peptide

[0046] 1) Preparation of streptavidin-coupled magnetic particle working solution

[0047] The streptavidin-coupled magnetic particle suspension was magnetically separated to remove the supernatant, and the HEPES buffer with a pH of 6.8 and a concentration of 0.2 mol / L was resuspended to a concentration of 1 mg / mL, and the buffer contained mass The percentage content is 2wt% bovine serum albumin, 0.02wt% Triton X-100, 0.05% sodium azide.

[0048] 2) Preparation of biotin-labeled MGF antibody

[0049] Take 1mg of MGF goat monoclonal antibody, add appropriate amount of PBS buffer (0.05M, pH7.0-7.6) to adjust the antibody concentration to 1mg / ml, and put it into a dialysis bag for dialysis, changing the dialysate every 3-4 hours, Replace 3-4 times, and transfer the antibody to a centrifuge tube or cryopreservation tube after dialysis;

[0050] Accurately...

Embodiment 3

[0066] Example 3 The preparation method of the chemiluminescent kit for detecting force growth factor and / or its E peptide

[0067] 1) Preparation of streptavidin-coupled magnetic particle working solution

[0068] The streptavidin-coupled magnetic particle suspension was magnetically separated to remove the supernatant, and resuspended to a concentration of 0.1 mg / mL in a phosphate buffer solution with a pH of 7.6 and a concentration of 0.01 mol / L. It contains 0.05% by weight of casein protein, 1% by weight of Brij35 and 0.5% of proclin 300 by weight.

[0069] 2) Preparation of biotin-labeled MGF-Ct24E antibody

[0070] Take 1mg of MGF-Ct24E sheep monoclonal antibody, add appropriate amount of PBS buffer (0.05M, pH7.0-7.6) to adjust the antibody concentration to 1mg / ml, and put it into a dialysis bag for dialysis, and change the dialysis every 3-4 hours solution, change it 3-4 times, and transfer the antibody to a centrifuge tube or cryopreservation tube after dialysis;

...

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Abstract

The invention relates to an adamantine chemiluminiscence kit for detecting a mechano-growth factor and the E peptide thereof and a manufacturing method of the kit. The prepared kit comprises streptavidin-coupled magnetic particle working fluid, biotin-labeled MGF and / or MGF-Ct24E antibody working fluid, alkaline phosphatase-labeled MGF and / or MGF-Ct24E antibody working fluid, MGF and MGF-Ct24E calibration product and / or quality control product working fluid, a chemiluminiscence substrate solution containing 3-(2-spiramantane)-4-methoxyl-4-(3-phosphorus oxygen acyl)-phenyl-1,2-dioxane disodiumsalt and cleaning fluid. A chemiluminescent system of the kit realizes chemiluminiscence through alkaline phosphatase / 3-(2-spiramantane)-4-methoxyl-4-(3-phosphorus oxygen acyl)-phenyl-1,2-dioxane disodium salt; by use of a streptavidin-biotin signal amplification system, the kit is high in detection sensitivity, wide in linear range and high in detection result repetitiveness, and can realize accurate quantification of the mechano-growth factor and the E peptide thereof.

Description

technical field [0001] The invention relates to an adamantane chemiluminescent kit for detecting force growth factor and / or its E peptide and its usage, belonging to the technical field of immune analysis. Background technique [0002] Force growth factor (MGF) was originally discovered in stretch-stimulated skeletal muscle. It is an important alloform produced by alternative splicing of the insulin-like growth factor-I (IGF-I) gene under force stimulation. Selection Splicing occurs at the site before exon 6 of the IGF-I gene, while MGF inserts 49 bp and 52 bp in the region of exon 5 in humans and rodents, respectively, resulting in a frameshift mutation in the translation of exon 6, A specific C-terminal 24 amino acid E peptide sequence (MGF-Ct24E) was generated. [0003] MGF and MGF-Ct24E can activate muscle satellite cells, promote tissue regeneration, promote osteoblast proliferation, and play a role in the response of osteoblasts or bone tissue to stress stimulation an...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/54326G01N2333/65G01N33/74G01N33/533
Inventor 李大军蔡淑娟徐霞飞涂策张金林
Owner SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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