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Preparation and application of human ICF45 (Interphase Cytoplasmic Foci Protein-45) monoclonal antibody

A monoclonal antibody, F6E4C12F11 technology, applied in anti-animal/human immunoglobulin, microbial-based methods, biochemical equipment and methods, etc., can solve the problems of cell proliferation and weakened adhesion

Active Publication Date: 2014-05-21
BEIJING NORMAL UNIVERSITY
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  • Claims
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Problems solved by technology

In the tumor cell model Hela, cell proliferation and attachment ability were found to be significantly weakened

Method used

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  • Preparation and application of human ICF45 (Interphase Cytoplasmic Foci Protein-45) monoclonal antibody
  • Preparation and application of human ICF45 (Interphase Cytoplasmic Foci Protein-45) monoclonal antibody
  • Preparation and application of human ICF45 (Interphase Cytoplasmic Foci Protein-45) monoclonal antibody

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Experimental program
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Embodiment 1

[0029] Embodiment 1, the preparation of monoclonal antibody

[0030] 1. Obtaining of hybridoma cell lines

[0031] 1. Preparation of ICF45 protein

[0032] The amino acid sequence of ICF45 protein is sequence 1 in the sequence listing, and the nucleotide sequence is sequence 2 in the sequence listing; the accession number of its full-length cDNA sequence on NCBI Genebank is AY463216.

[0033] Design specific primers with BamH I restriction sites at both ends:

[0034] Upstream primer 5'-CGC GGATCC TGGGGCGCCTGTAAA-3';

[0035] Downstream primer 5'-CGC GGATCC CGCAAAAGGGTCAG-5';

[0036] The nucleotide sequence (sequence 1) of the artificially synthesized ICF45 protein was used as a template, and the above-mentioned upstream primer and downstream primer were used for PCR amplification to obtain a PCR product.

[0037] The PCR product was digested with BamHI, and the recovered digested product was mixed with pGEX-PUC-3T (this plasmid was constructed by pGEX-2t, pGEX-2t was...

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Abstract

The invention discloses preparation and application of a human ICF45 (Interphase Cytoplasmic Foci Protein-45) monoclonal antibody and provides a mus musculus hybridoma F6E4C12F11 having a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.8456 as well as a monoclonal antibody generated by the mus musculus hybridoma F6E4C12F11 having the preservation number of CGMCC No.8456. An experiment shows that the mus musculus hybridoma F6E4C12F11 is obtained in a screening manner and is subjected to biological characteristic identification, so that the influence of the protein level of the ICF45 to a normal physiological process of a cell is further probed, the basis and the foundation for exerting corresponding action relation in diseases, tumors and allergic reactions and deeply researching the functions of the mus musculus hybridoma are provided and laid, and potential drug proteins for suppressing the corresponding diseases are prepared.

Description

technical field [0001] The present invention relates to the preparation and application of an interphase cytoplasmic focus protein-45 (Interphase Cytoplasmic Foci Protein-45, ICF45) monoclonal antibody, in particular to the preparation and application of a human ICF45 monoclonal antibody. Background technique [0002] ICF45 protein is a new cytoplasmic protein discovered, its expression has high cell cycle phase specificity, and plays an important role in cell cycle regulation. Previous studies have shown that ICF45 is localized in the cytoplasm during the interphase of the cell, forms 1-2 specific point-like focuses near the nuclear membrane, and co-localizes with the centrosome in a cell cycle phase-specific manner, while in the mitotic phase of the cell disappear. [0003] Numerous studies have suggested that the occurrence of tumors is related to the amplification of the centrosome. Abnormal duplication of centrosomes, such as increased number of centrosomes, can cause...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/18G01N33/68C12R1/91
Inventor 何大澄商瑜伊力亚尔王珊珊肖雪媛
Owner BEIJING NORMAL UNIVERSITY
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