Rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody and preparation method thereof

A ubiquitin-conjugating enzyme and polyclonal antibody technology, applied in the field of bioengineering, can solve the problems of limitations, lack of material basis and technical support, and no production method of cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody, etc., to achieve specificity Good, high-purity effect

Inactive Publication Date: 2015-11-04
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Polyclonal antibody technology is an important part of the study of protein function and clinical detection, but in the current prior art, the research on the cucumber ubiquitin-conjugating enzyme (E2) gene is not yet mature, and the polyclonal antibody to the cucumber ubiquitin-conjugating enzyme CsE2 protein There have been no related reports, let alone a specific preparation method for the polyclonal antibody to the cucumber ubiquitin-conjugating enzyme CsE2 protein. The material basis and technical support for chemical plant detection, variety purity and molecular biology research

Method used

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  • Rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody and preparation method thereof
  • Rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody and preparation method thereof
  • Rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody and preparation method thereof

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Embodiment 1

[0030] A cucumber ubiquitin-conjugating enzyme CsE2 protein used for preparing a rabbit polyclonal antibody against cucumber ubiquitin-conjugating enzyme CsE2 protein according to an embodiment of the present invention, the amino acid sequence of the protein is shown in SEQ ID NO:1.

[0031] The embodiment of the present invention provides a rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody, which is produced by rabbits with the cucumber ubiquitin-conjugating enzyme CsE2 protein as shown in SEQ ID NO:1 as an immunogen polyclonal antibody. The specific structure of the rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody structure is as follows image 3 shown.

[0032] On the other hand, the embodiment of the present invention also provides a method for preparing a rabbit polyclonal antibody against cucumber ubiquitin-conjugating enzyme CsE2 protein, comprising the following steps:

[0033] Step 1 purifying the cr...

Embodiment 2

[0043] 1. Purification and purity analysis of antigenic protein

[0044] Cucumber ubiquitin-conjugating enzyme CsE2 protein was extracted by cucumber shoot tip, and then the crude product of antigenic protein was obtained by purification and two-dimensional electrophoresis, and then the crude product of antigenic protein was used on GE’s Protein G affinity column (HiTrap Protein G HP 17-0404-01) Purification and purity analysis were carried out so that the purity of cucumber ubiquitin-conjugating enzyme CsE2 protein reached 95.8%.

[0045] 2. Immunization of experimental animals

[0046]One healthy male rabbit was selected, and the negative serum was extracted from the ear vein of the rabbit as the serum of the control group. Select 200 μg of cucumber ubiquitin-conjugating enzyme CsE2 protein purified in step 1, dissolve and dilute it with 200 μL of normal saline, add an equal volume of Freund’s adjuvant to emulsify cucumber ubiquitin-conjugating enzyme CsE2 protein (initial ...

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Abstract

The present invention provides a rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody and a preparation method thereof. The method comprises: extracting ubiquitin-conjugating enzyme CsE2 protein from cucumber stem apex; carrying out two-dimensional electrophoresis, and purifying to obtain antigen; injecting the antigen into rabbits to immunize; and drawing serum, and purifying to obtain the final rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody. According to the present invention, The prepared rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody has characteristics of good specificity and high purity, can be subjected to specific binding reaction with cell endogenous cucumber ubiquitin-conjugating enzyme CsE2 protein, can meet requirements of immunohistochemistry, immunoblotting and other experiments, and can be used for establishment of in vitro immunoassays; and the preparation method provides the important tool for the deep research of the biological functions of the cucumber ubiquitin-conjugating enzyme CsE2 protein and the identification of the cucumber plant dwarf, and has important theoretical significance and practical application values.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a rabbit anti-cucumber ubiquitin-conjugating enzyme CsE2 protein polyclonal antibody and a preparation method thereof. Background technique [0002] An antigen is usually composed of multiple antigenic determinants. One antigenic determinant stimulates the body, and the antibody produced by a B lymphocyte receiving the antigen is called a monoclonal antibody. The body is stimulated by a variety of antigenic determinants, and correspondingly various monoclonal antibodies are produced. These monoclonal antibodies are mixed together to form polyclonal antibodies, and the antibodies produced in the body are polyclonal antibodies; In addition to diversity, the same type of antigenic determinant can also stimulate the body to produce five types of antibodies including IgG, IgM, IgA, IgE and IgD. Polyclonal antibody is a group of immunoglobulins secreted by plasma cells of the o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/00C07K16/40C07K16/06C07K1/22
CPCC07K16/06C07K16/40C12N9/93C12Y603/02019
Inventor 辛明
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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