Porcine P21 protein antibody as well as preparation method and application thereof

A P21, protein technology, applied in the direction of serum immunoglobulin, anti-animal/human immunoglobulin, material test products, etc. Strong demand for antibodies and other issues, to achieve the effect of broad application prospects

Inactive Publication Date: 2014-06-25
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are many studies on the p21 gene in humans and mice, and many antibodies against the p21 gene of humans and mice have been produced, but the preparation of antibodies against the p21 gene of pigs at home and abroad is very rare, because pigs are used as experiments. In the research of materials, it is often encountered that no suitable antibody can be found when detecting the expression of a certain protein; however, the commercially available human and mouse p21 antibodies cannot detect porcine p21 protein well, which makes many The research on pigs cannot be carried out comprehensively; with the in-depth development of pig molecular genetic breeding work, the demand for pig-specific antibodies is becoming more and more intense

Method used

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  • Porcine P21 protein antibody as well as preparation method and application thereof
  • Porcine P21 protein antibody as well as preparation method and application thereof
  • Porcine P21 protein antibody as well as preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0033] The preparation of pig p21 protein antibody, the technical route is as follows figure 1 , Including the following steps:

[0034] S1. Cloning of pig P21 gene

[0035] In the present invention, the total RNA of porcine skeletal muscle satellite cells is first extracted, cDNA is obtained by reverse transcription, and the porcine P21 gene is cloned by using the cDNA as a template. It includes the following steps:

[0036] S11. Extraction of total RNA: For extraction of total RNA from porcine skeletal muscle satellite cells, refer to the instruction manual of RNAiso Plus Total RNA Extraction Reagent from TAKARA.

[0037] S12. Reverse transcription of total RNA: The reverse transcription of the extracted total RNA is performed according to the instructions of TIANGEN's TIANScript cDNA first-strand synthesis kit. The specific steps are as follows:

[0038] Add the following reaction mixture to a nuclease-free centrifuge tube in an ice bath:

[0039]

[0040] Reaction conditions: After ...

Embodiment 2

[0118] The specificity of the anti-pig P21 antibody prepared in Example 1 of the present invention is compared with the commercially available anti-mouse P21 antibody. The specific method is: comparing the anti-pig P21 antibody prepared by the present invention with the commercially available anti-mouse P21 antibody by Western blot. The ability of P21 antibody to recognize P21 protein of pigs and mice, such as Picture 11 Shown from Picture 11 It can be seen that the anti-pig P21 antibody prepared in Example 1 of the present invention can well recognize the P21 protein of pigs and mice; while the anti-mouse P21 antibody purchased on the market can only recognize the P21 protein of mice to a limited extent. The P21 protein of pigs is basically not recognized. It can be seen that the anti-pig P21 protein antibody prepared by the present invention has specificity to the pig P21 protein.

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Abstract

The invention discloses a porcine P21 protein antibody as well as a preparation method and an application thereof. Porcine P21 protein obtained through expression and purification is used for preparing an immunogen to immunize a purebred New Zealand rabbit so as to prepare a rabbit anti-pig P21 protein antibody; and the prepared rabbit anti-pig P21 protein antibody is capable of well recognizing the P21 protein of a pig and a mouse, while a commercially available anti-human P21 antibody is only capable of limitedly recognizing the P21 protein of the mouse. The porcine P21 protein antibody prepared in the invention fills up the blank in deficiency of a porcine specific antibody and provides materials for a scientific research of porcine genetic breeding.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to a pig P21 protein antibody and its preparation method and application. Background technique [0002] In animal genomes, p21, cdkl, p53, checkl, helb, atm and other genes are the main genes that maintain the genetic stability of animals, and their stable expression is of great significance to the growth, maintenance and differentiation of cells. [0003] The p21 gene is an important member of the cyclin-dependent kinase inhibitory family discovered in recent years, and the protein encoded by it, p21 protein, is an important cell cycle regulatory protein. Participate in the process of cell growth, differentiation, aging and death; p21 protein is also closely related to the occurrence of tumors, and plays an important role in the physiological and pathological processes of cells: studies have shown that p21 can form a complex with cyclin D / CDK to enable T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K16/06G01N33/68
Inventor 王翀徐建吴珍芳张守全余丽明邹潭焦义仁段军莉史中元蒋承凤
Owner SOUTH CHINA AGRI UNIV
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