Preparation method and application of polyclonal antibody for specifically recognizing protein USP42

A polyclonal antibody and protein recognition technology, applied in biochemical equipment and methods, anti-enzyme immunoglobulin, and the use of vectors to introduce foreign genetic material, etc., to achieve the effects of high expression yield, robust physique, and easy access

Inactive Publication Date: 2019-12-03
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, in the prior art, the relevant research on the preparation method of the protein USP42 polyclonal antibody is still in a blank stage

Method used

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  • Preparation method and application of polyclonal antibody for specifically recognizing protein USP42
  • Preparation method and application of polyclonal antibody for specifically recognizing protein USP42
  • Preparation method and application of polyclonal antibody for specifically recognizing protein USP42

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Embodiment

[0032] The polyclonal antibody is a mouse IgG2a subtype polyclonal antibody.

[0033] The preparation method of the above-mentioned polyclonal antibody specifically recognizing protein USP42 comprises the following steps:

[0034] (1) Obtain the USP42 protein fragment; select the target fragment by antigenic epitope analysis for recombinant expression of protein USP42, the gene sequence of the target fragment is shown in SEQ ID No.1 in the sequence listing, and its corresponding amino acid sequence is shown in sequence As shown in SEQ ID No.4 in the list, BamH1, NotI restriction sites and homology arms were introduced into the upstream and downstream primers respectively;

[0035] An upstream primer, the specific sequence of which is shown in SEQ ID No.2 in the sequence listing;

[0036] A downstream primer, the specific sequence of which is shown in SEQ ID No.3 in the sequence listing;

[0037] (2) extract the normal testicular tissue mRNA of the mouse, and use it as a temp...

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Abstract

The invention relates to the technical field of bioscience and detection reagents. The invention specifically discloses a preparation method and application of a polyclonal antibody for specifically recognizing protein USP42. The method comprises the following steps: (1), protein USP42 antigenic epitope analysis: selecting a target fragment as recombinant expression of protein USP42 through the antigenic epitope analysis, wherein the gene sequence of the target fragment is shown as SEQ ID N0.1 in a sequence table; and respectively introducing BamH1 and Not I restriction enzyme cutting sites into an upstream primer and a downstream primer. According to the invention, the preparation of the polyclonal antibody of USP42 is realized.

Description

technical field [0001] The invention relates to the technical fields of biological science and detection reagents. Background technique [0002] Deubiquitinating enzyme USP42 (Ubiquitin specific peptidase 42) is a member of the USP family of deubiquitinating enzymes (DUBs). Ubiquitination is the labeling effect of ubiquitin, which is catalyzed by three enzymes. Ubiquitin is a protein composed of 76 amino acids that is widely found in eukaryotic cells. The protein degradation pathway mediated by ubiquitin is called "ubiquitin-proteasome pathway". The protein to be degraded is released as a degradation product after unfolding, opening, and hydrolysis. Ubiquitination is a reversible process. Ubiquitination is a cascade of enzymes that cause ubiquitin to mark selected proteins. Deubiquitination separates ubiquitin from proteins by deubiquitinases. The processes of deubiquitination and ubiquitination constitute protein homeostasis in vivo. At present, research on deubiquitin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/40C12N15/70G01N33/573C12N9/48
CPCC07K16/40C12N9/485C12N15/70C12Y304/19012G01N33/573G01N2333/948
Inventor 刘明兮刘思钰吴佳艳
Owner NANJING MEDICAL UNIV
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