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Antigen protein of anti-bridge molecule 1-s antibody, and rabbit polyclonal antibody and application thereof

A technology of polyclonal antibody and antigenic protein, applied in the field of biomedicine

Active Publication Date: 2015-03-11
TIANJIN MEDICAL UNIV CANCER INST & HOSPITAL
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AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the problem that currently available ITSN1-S antibodies have poor specificity and low titer ratio, making it difficult to apply to the detection of ITSN1-S protein at the cell level and tissue level The problem is to provide an antigenic protein of anti-bridge molecule 1-s antibody and its rabbit polyclonal antibody and use

Method used

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  • Antigen protein of anti-bridge molecule 1-s antibody, and rabbit polyclonal antibody and application thereof
  • Antigen protein of anti-bridge molecule 1-s antibody, and rabbit polyclonal antibody and application thereof
  • Antigen protein of anti-bridge molecule 1-s antibody, and rabbit polyclonal antibody and application thereof

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Embodiment Construction

[0021] 1. Experimental materials:

[0022] The human malignant glioblastoma cell line LN-229 cell line was purchased from the American ATCC cell bank;

[0023] The tissue samples of human breast cancer patients are derived from the pathological paraffin sections of patients undergoing tumor resection in the Department of Breast Surgery;

[0024] Antigen: Human ITSN1-S protein sequence (NM_001001132) was used to select protein immune fragments to obtain specific antigenic fragments targeting human ITSN1-S, qpvaissapafgmggias, artificially synthesized peptides, and the purity of the synthetic peptides was greater than or equal to 95%.

[0025] Phosphate buffer saline (PBS): 8 grams of sodium chloride, 0.2 grams of potassium chloride, 3.63 grams of dibasic sodium phosphate, 0.24 grams of potassium dihydrogen phosphate, dissolved in 1 liter of pure water, pH 7.4.

[0026] TBS buffer solution: 10mmol / L Tris containing 0.9% NaCl, adjust the pH to 7.4 with HCl.

[0027] Elution buf...

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Abstract

The invention discloses an antigen protein of an anti-bridge molecule 1-s antibody, and a rabbit polyclonal antibody and application thereof, belonging to an antigen / antibody-containing pharmaceutical preparation. The sequence of the specific antigen protein of anti-human bridge molecule 1-s antibody is qpvaissapafgmggias. The rabbit polyclonal antibody is prepared by the following steps: immunizing a rabbit with the specific antigen of the anti-human bridge molecule 1-s antibody to obtain an antiserum, emulsifying to obtain an antigen solution, injecting into an adult rabbit, and collecting antibody-containing blood, wherein the clear solution is the antiserum; and balancing an antigen polypeptide affine column by using a precooled TBS buffer solution, adding sodium azide, carrying out centrifugal filtration, degreasing, washing the column, and purifying to obtain the anti-human bridge molecule 1-s rabbit polyclonal antibody. The polyclonal antibody has obvious better expression than the commercial on-sale antibody on the cellular level and tissue level. The polyclonal antibody solves the problems of poor specificity and low potency ratio in the existing on-sale commodity anti-human ITSN 1-S antibody.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an antigenic protein of a bridge molecule 1-s antibody, a rabbit polyclonal antibody and its application. Background technique [0002] Bridge molecule 1 (intersectin1 or ITSN1) is a highly conserved protein in the evolution process, and there are two spliced ​​forms of ITSN1-L and ITSN1-S in the human body. In the central nervous system, ITSN1-L is mainly distributed in neurons, while ITSN1-S is mainly expressed in glial cells. The two splice forms share several common protein domains. ITSN1-L protein is associated with a variety of neurodegenerative diseases, such as: congenital stupidity; Alzheimer's disease (AD) and Huntington's disease, etc. The occurrence of congenital stupidity is related to the disorder of vesicle transport regulated by ITSN1-L, while the occurrence of AD and Huntington's disease is related to the neurotoxicity of polyglutamine promoted by ITSN1-L. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K16/18C07K16/06G01N33/68
CPCC07K14/47C07K16/06C07K16/18G01N33/68
Inventor 马勇杰应国光谷峰付丽王海龙
Owner TIANJIN MEDICAL UNIV CANCER INST & HOSPITAL
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