A kind of monoclonal antibody of RNA polymerase II transcriptional subunit 37e mediator and its application

An RNA polymerase, antibody technology, applied in anti-enzyme immunoglobulins, instruments, peptides, etc., can solve problems such as lack of antibody tools

Active Publication Date: 2021-07-06
艾比玛特医药科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no targeting antibody specifically targeting the RNA polymerase II transcription subunit 37e mediator in the study of the toxic carcinogen PAHs in crops, and there is a lack of effective antibody tools for this research field at the proteomics level

Method used

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  • A kind of monoclonal antibody of RNA polymerase II transcriptional subunit 37e mediator and its application
  • A kind of monoclonal antibody of RNA polymerase II transcriptional subunit 37e mediator and its application
  • A kind of monoclonal antibody of RNA polymerase II transcriptional subunit 37e mediator and its application

Examples

Experimental program
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preparation example Construction

[0124] Preparation of monoclonal antibodies

[0125] Antibodies of the present invention can be prepared by various techniques known to those skilled in the art. For example, an antigen of the invention may be administered to an animal to induce the production of monoclonal antibodies. For monoclonal antibodies, hybridoma technology can be used to prepare (see Kohler et al., Nature 256; 495, 1975; Kohler et al., Eur.J.Immunol.6:511, 1976; Kohler et al., Eur.J.Immunol. 6:292,1976; Hammerling et al., In Monoclonal Antibodies and T Cell Hybridomas, Elsevier, N.Y., 1981) or can be prepared by recombinant DNA methods (US Patent No. 4,816,567).

[0126]Representative myeloma cells are those that fuse efficiently, support stable high-level production of antibody by selected antibody-producing cells, and are sensitive to culture medium (HAT medium matrix), including myeloma cell lines, such as murine Myeloma cell lines, including those derived from MOPC-21 and MPC-11 mouse tumors (a...

Embodiment 1

[0152] Example 1 Elisa (immunoenzyme-linked) pairing verification of antibody and antigenic polypeptide

[0153] Coat the 96-well ELISA plate with ascitic fluid antibody to be paired, incubate, wash and block overnight with skim milk, wash with PBS, and store at 4°C until use. Antigen peptides were incubated, washed with PBS, and controls were set at the same time. HRP-labeled detection antibody was added to the previously incubated ELISA plate. TMB color reaction, microplate reader reading. The titers of the 8 cell lines obtained by our screening are shown in Table 1:

[0154] Table 1 Titers of 8 cell lines

[0155]

Embodiment 2

[0156] Endogenous western blotting (WB) verification of embodiment 2 antibody

[0157] Use the whole protein lysate of pear, and the antibody dilution concentration is 1:1000, 1:2000 and 1:5000 for WB verification. experiment result shows( figure 2 ), anti-MED37E (clone 3F12) can specifically recognize the 69kd band in the WB verification, which is consistent with the expected size.

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Abstract

The invention provides a monoclonal antibody of RNA polymerase II transcription subunit 37e mediator and application thereof. Experimental results show that the antibody of the present invention can be applied to antibody chip reaction, immunoprecipitation, Western Blotting and detection of the content of toxic and carcinogenic polycyclic aromatic hydrocarbons (PAHs) in plant roots.

Description

technical field [0001] The invention belongs to the field of biomedicine, specifically, the invention relates to a monoclonal antibody of RNA polymerase II transcription subunit 37e mediator and application thereof. Background technique [0002] The RNA polymerase II transcriptional subunit 37e mediator is a component of a coactivator encoded by the MED37E gene that is involved in nearly all RNA polymerase II-dependent gene-regulated transcription. This protein is a molecular chaperone, belongs to the heat shock protein 70 family and widely exists in plants, especially in crop roots. [0003] Polycyclic aromatic hydrocarbons (PAHs) are potentially carcinogenic and toxic to humans, and humans can become ill through ingestion of contaminated food. At present, there is no targeting antibody specifically targeting the RNA polymerase II transcription subunit 37e mediator in the study of toxic carcinogens PAHs in crops, and there is a lack of effective antibody tools for this res...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/40G01N33/577G01N33/53
CPCC07K16/40C07K2317/565G01N33/5308G01N33/577
Inventor 翁炜宁张玉娜孟逊
Owner 艾比玛特医药科技(上海)有限公司
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