Preparation method and application of bovine VCAM-1 polyclonal antibody

A VCAM-1, polyclonal antibody technology, applied in chemical instruments and methods, anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve biological functions Limitation, low homology, etc., to avoid false positives and ensure purity and quality

Inactive Publication Date: 2016-07-27
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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Problems solved by technology

In order to study the immune mechanism and function of VCAM-1 in dairy cow mastitis, a high-quality purified protein and efficient and specific VCAM-1 antibody are essential, however, the difference between bovine and mouse VCAM-1 The homology of VCAM-1 is extremely low, and the research on the biological function of bovine VCAM-1 is limited due to the lack of commercial antibodies to bovine VCAM-1

Method used

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  • Preparation method and application of bovine VCAM-1 polyclonal antibody
  • Preparation method and application of bovine VCAM-1 polyclonal antibody
  • Preparation method and application of bovine VCAM-1 polyclonal antibody

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Embodiment Construction

[0036] Below in conjunction with experiment the present invention is further described:

[0037] The preparation method of bovine source VCAM-1 polyclonal antibody comprises the following steps:

[0038] 1. Intestines of healthy dairy cows containing Peyer's node tissue were routinely stored in a -80°C low-temperature refrigerator in the laboratory. Three Wistar rats, weighing 180-220 g, were purchased from the Experimental Animal Center of Jilin University School of Basic Medicine. RNA extraction (Trizol method, carried out according to the molecular cloning experiment guideline), extracting RNA from the intestinal tube containing Peyer's node tissue in healthy dairy cows.

[0039] 2. See Table 1 and Table 2 for the reverse transcription PCR (according to the instructions) system and reaction conditions.

[0040] Table 1 reverse transcription PCR reaction system

[0041]

[0042]

[0043] Table 2 Carry out the reverse transcription reaction according to the following c...

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Abstract

The invention belongs to a preparation method and application of a bovine VCAM-1 polyclonal antibody. The method comprises the following steps of (1) RNA (ribonucleic acid) extraction; (2) reverse transcription PCR (polymerase chain reaction); (3) primer design and synthesis; (4) gene fragment amplification; (5) gene clone; (6) prokaryotic expression vector building; (7) expression and identification of DNA (deoxyribonucleic acid) recombinant plasmid in escherichia coli; (8) recombinant protein purification; (9) protein blotting; (10) recombinant protein dialysis and concentration determination; (11) immunizing antigen preparation; (12) immunizing and blood collection; (13) serum separation. The polyclonal antibody provided by the invention can be used for identifying the bovine recombinant VCAM-1 protein and natural recombinant VCAM-1 protein; the valence of the VCAM-1 polyclonal antibody is high; the specificity is high. The antibody particularly aims at the bovine natural VCAM-1 protein.

Description

technical field [0001] The invention relates to a preparation method and application of bovine VCAM-1 polyclonal antibody. Background technique [0002] VCAM-1 was first cloned by Osbom et al. in 1989 using eukaryotic gene expression methods. VCAM-1 is mainly expressed on the surface of endothelial cells, epithelial cells, macrophages, and dendritic cells, and is involved in the adhesion and migration of immune cells. . In order to study the immune mechanism and function of VCAM-1 in dairy cow mastitis, a high-quality purified protein and efficient and specific VCAM-1 antibody are essential, however, the difference between bovine and mouse VCAM-1 Due to the lack of commercial bovine VCAM-1 antibody, the research on the biological function of bovine VCAM-1 is limited. Contents of the invention [0003] The present invention provides a preparation method and application of a bovine VCAM-1 polyclonal antibody. The polyclonal antibody of the present invention can be used to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C07K16/06C12N15/70G01N33/68
CPCC07K16/2836C07K2317/10
Inventor 常巧呈陈媛媛张乃生杨正涛罗英花
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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