Preparation method of polyclonal antibody for specific recognition of protein MEX3D and application thereof

A polyclonal antibody and protein recognition technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, applications, etc., to achieve the effects of strong adaptability, strong physique, and strong reproductive ability

Inactive Publication Date: 2019-11-05
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, the relevant research on the preparation method of protein MEX3D polyclonal antibody in the prior art is still in a blank stage

Method used

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  • Preparation method of polyclonal antibody for specific recognition of protein MEX3D and application thereof
  • Preparation method of polyclonal antibody for specific recognition of protein MEX3D and application thereof
  • Preparation method of polyclonal antibody for specific recognition of protein MEX3D and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0028] The polyclonal antibody is a mouse IgG2a subtype polyclonal antibody.

[0029] The preparation method of the above-mentioned polyclonal antibody specifically recognizing protein MEX3D comprises the following steps:

[0030] (1) Obtain the MEX3D protein fragment; select the target fragment by antigenic epitope analysis for recombinant expression of the protein MEX3D, the gene sequence of the target fragment is shown in SEQ ID No.1 in the sequence table, and its corresponding amino acid sequence is shown in sequence As shown in SEQ ID No.4 in the list, the upstream and downstream primers introduce EcoRI, Hind III restriction sites and their homology arms respectively; the upstream primers are as shown in SEQ ID No.2 and the downstream primers are as shown in SEQ ID No. 3.

[0031] (2) extract the normal testicular tissue mRNA of the mouse, and use it as a template to carry out reverse transcription PCR, synthesize the first strand of cDNA and then use it as a template to...

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Abstract

The invention relates to the technical field of biological science and detection reagent, and in particular, relates to a preparation method of a polyclonal antibody for specific recognition of a protein MEX3D and an application thereof, wherein the preparation method includes the following steps: protein MEX3D antigen epitope analysis: a target fragment is selected for recombinant expression of the protein MEX3D through antigen epitope analysis, a gene sequence of the target fragment is shown in SEQ ID NO.1 in a sequence table, and an EcoRI and Hind III enzyme digestion site and homologous arms and the like are introduced into upstream and downstream primers respectively. The polyclonal antibody of MEX3D is prepared.

Description

technical field [0001] The invention relates to the technical fields of biological science and detection reagents. Background technique [0002] The protein MEX3D is a protein encoded by the gene MEX3D (mex-3RNA binding family member D). [0003] Mex-3 protein is a translation regulator. In the early embryo of Caenorhabditis elegans, Mex-3 can recognize blastocyst and maintain cell pluripotency (reference: Buchet-Poyau, K. and J. Courchet, et al. (2007). "Identification and characterization of human Mex-3proteins, a novel family of evolutionarily conserved RNA-binding proteins differentially localized to processing bodies." Nucleic Acids Res 35(4):1289-300). [0004] Human homologous protein Mex-3 is a phosphorylated protein, including MEX-3A~3D four proteins. Among them, MEX3D is widely expressed in various human tissues (PMID 24309898), and is a class of conserved RNA-binding proteins. [0005] Studies have shown that MEX3D has an important regulatory effect on the expr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K14/47C12N15/12C12N15/70G01N33/68
CPCC07K14/47C07K16/18C12N15/70G01N33/68G01N2333/47
Inventor 刘明兮张金涛吴佳艳
Owner NANJING MEDICAL UNIV
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