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Detection method of myocardial cell M3 receptor

A technology of cardiomyocytes and detection methods, applied in the field of cardiomyocyte M3 receptors, can solve the problems of cumbersome detection methods of cardiomyocytes M3 receptors, expensive detection reagents or instruments, and long time-consuming, so as to achieve easy routine and large-scale use , low cost, and the effect of improving accuracy

Pending Publication Date: 2021-11-02
刘艳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem that the existing detection method of cardiomyocyte M3 receptor is cumbersome, takes too long, has low accuracy, and the detection reagents or instruments used are expensive

Method used

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  • Detection method of myocardial cell M3 receptor
  • Detection method of myocardial cell M3 receptor
  • Detection method of myocardial cell M3 receptor

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Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0032] Specific embodiment one: the present invention provides a kind of detection method of cardiomyocyte M3 receptor, comprises the following steps:

[0033] S1 peptide synthesis: take 5-10mL samples to synthesize peptides according to the second loop functional epitope peptide sequence of the cardiomyocyte M3 receptor extracellular peptide sequence, the second loop functional epitope peptide sequence is CLFWQYFVGKRTVPPGEC at positions 205-220, Purify the synthesized peptide to 93%-98% by reverse-phase high-performance liquid chromatography to obtain a purified sample;

[0034] S2 ELISA plate coating: choose 50mM, pH9.6 carbonate coating buffer to dilute the purified sample to obtain 10μg·mL -1 The diluted sample solution, inject 100 μL of the diluted sample solution into each well of the microtiter plate for coating, and place it at 4°C for 12h-18h;

[0035] S3 blocking: Remove the coating solution from the coated wells of the microtiter plate, wash with PBST 4-6 times, 3 ...

specific Embodiment approach 3

[0044] Embodiment 3: The enzyme plate described in S2 is a 96-well medium-binding enzyme plate. The selection of medium-binding enzyme plate has the characteristics of high detection specificity and sensitivity, and low blank hole value. The other combinations and connections of this embodiment are the same as those of the second embodiment.

[0045] Embodiment 4: The diluted horseradish peroxidase-labeled goat anti-human IgG solution described in S4 is obtained by diluting horseradish peroxidase-labeled goat anti-human IgG with water at a mass ratio of 1:1000. The other combinations and connections of this embodiment are the same as those of the third embodiment.

[0046] Embodiment 5: The chromogenic agent described in S5 is a TMB substrate chromogenic agent or an OPD substrate chromogenic agent. The TMB substrate chromogenic reagent has high stability and lower sensitivity than the OPD substrate chromogenic reagent, and the OPD substrate chromogenic reagent has high sensi...

specific Embodiment approach 7

[0048] Embodiment 7: When the chromogenic agent is a TMB substrate chromogenic agent, use a microplate reader A490 to measure in S6; when the chromogenic agent is an OPD substrate chromogenic agent, use a microplate reader A450 to measure in S6. The other combinations and connections of this embodiment are the same as those of Embodiment 6.

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Abstract

The invention provides a detection method of a myocardial cell M3 receptor, and belongs to the technical field of myocardial cell M3 receptors. The problems that an existing myocardial cell M3 receptor detection method is tedious, consumed time is too long, the accuracy rate is low, and used detection reagents or instruments are high in price are solved. The method comprises the following steps: synthesizing a second ring functional epitope peptide fragment sequence outside a myocardial cell M3 recipient cell into a peptide fragment, purifying, dissolving with a carbonate coating buffer solution, coating with an elisa plate, adding 300 [mu] L of PBS containing milk with a volume fraction of 5% into each pore, sealing, adding an elisa sample, developing, and determining an OD value through a microplate reader. As the catalysis frequency of the enzyme is high, the detection method adopted by the invention can amplify the reaction effect and effectively improve the accuracy of the measurement result; the method has the advantages that complex detection links are not needed, used reagents and microplate reader are common reagents and instruments, the method is simple, materials are easy to obtain, cost is low, and conventional and large-scale use is easy; and meanwhile, the method is sensitive and efficient.

Description

technical field [0001] The invention relates to the technical field of cardiomyocyte M3 receptors, in particular to a method for detecting cardiomyocyte M3 receptors. Background technique [0002] Autoantibodies are antibodies directed against one's own tissues, organs, cells and cellular components. The normal immune response of healthy individuals is a protective defense that removes foreign antigens from the body. Usually, the ubiquitous autologous and antigenic reactive organism can tolerate it. However, the lack of selection in this process results in self-tissue damage, which leads to the continuous production of new self-antigens, which in turn induces a persistent immune response. In recent years, the role of immune system abnormalities in the occurrence and development of heart failure has gradually been paid attention to, such as autoantibodies to G protein-coupled receptors such as β1, β2 adrenergic receptors and M2-type acetylcholine receptors. The production ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543G01N33/68
CPCG01N33/535G01N33/54306G01N33/68
Inventor 刘艳蒋雅楠郭悦平柏冰雪王姝高亚男
Owner 刘艳
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