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RAC1 protein monoclonal antibody as well as preparation method and application thereof

A monoclonal antibody, RAC1 technology, applied in the field of biological immunology, can solve the problems of poor specificity of polyclonal antibodies, and achieve the effect of strong specificity, convenient operation, and high synthesis efficiency

Pending Publication Date: 2022-06-07
SHAOXING SHOUREN MEDICAL HEALTH & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of this application is to provide a RAC1 protein monoclonal antibody and its preparation method and application, aiming to solve the problem of poor specificity of the RAC1 protein polyclonal antibody in the prior art, which is not conducive to accurate and rapid detection of RAC1 protein

Method used

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Experimental program
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Effect test

preparation example Construction

[0031] The second aspect of the present application provides a method for preparing a RAC1 protein monoclonal antibody, the preparation method of the RAC1 protein monoclonal antibody comprises the following steps:

[0032] S01. Dissolve RAC1 protein in buffer to obtain RAC1 protein solution, RAC1 protein solution for immune mice, obtain immune mice, and immune serum;

[0033] S02. Splenocytes and myeloma cells of immune mice were cultured separately, and cell fusion treatment was performed, and indirect ELISA and indirect competition ELISA were used for screening to obtain hybridoma cell lines.

[0034]S03. Cell cloning of hybridoma cell lines to obtain strongly positive monoclonal cells;

[0035] S04. Strong positive monoclonal cells were inoculated into mice, and RAC1 protein monoclonal antibodies were prepared by in vivo induced ascites method.

[0036] Embodiment of the present application provided in the second aspect of the preparation method of RAC1 protein monoclonal antib...

Embodiment 1

[0065] RAC1 protein monoclonal antibody and preparation method thereof

[0066] Among them, the sequence of the RAC1 protein monoclonal antibody includes: the amino acid sequence SEQ ID NO.1 of the heavy chain variable region of the RAC1 protein monoclonal antibody sequence and the amino acid sequence SEQID NO.2 of the light chain variable region of the RAC1 protein monoclonal antibody sequence.

[0067] The preparation method of the RAC1 protein monoclonal antibody is as follows:

[0068] (1) Preparation of antigens: Weigh 100mg RAC1 protein dissolved in 1.8mL PBS, adjust pH to 7.4, volume to 2 ml, and prepare 50mg / ml RAC1 protein solution.

[0069] (2) Animal immunity and titer determination

[0070]Take a number of 6 to 8 weeks old healthy BALB / c mice numbered, take 50mg / ml antigen solution, mix well with equal volume of complete Freund's adjuvant, after full emulsification, press 100 μg / only immune mice for primary immunization, and then strengthen immunity every 2 weeks to 4 ...

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Abstract

The invention relates to the technical field of biological immunology, in particular to an RAC1 protein monoclonal antibody as well as a preparation method and application thereof, and provides the RAC1 protein monoclonal antibody, and the sequence of the RAC1 protein monoclonal antibody is as follows: the amino acid sequence of a heavy chain variable region of the RAC1 protein monoclonal antibody sequence is SEQ ID NO.1, and the amino acid sequence of a heavy chain variable region of the RAC1 protein monoclonal antibody sequence is SEQ ID NO.2; the amino acid sequence of the light chain variable region of the RAC1 protein monoclonal antibody sequence is SEQID NO.2; the RAC1 protein monoclonal antibody provided by the invention has strong specificity, can accurately and quickly detect RAC1 protein, is specifically combined with RAC1 protein, ensures that the RAC1 protein monoclonal antibody has no cross reaction with other proteins in an RHO protein family, and is beneficial to wide application in immunological detection.

Description

Technical field [0001] The present application belongs to the field of biological immunology technology, in particular to a RAC1 protein monoclonal antibody and its preparation method and application. Background [0002] Rac 1 is a member of the Rho family. The Rho family plays a role as a "molecular switch" in some of the basic functions of cells, involved in the movement of cells, actin recombination, malignant transformation of tumors, invasion and metastasis, regulation of transcription factors, apoptosis of cells, and angiogenesis of tumors. The protein encoded by the Rac1 gene is GTPase, which belongs to the RAS superfamily of small GTP-binding proteins; It plays an important role in cell motility and adhesion, cell proliferation, differentiation and apoptosis, tumor invasion and metastasis, and immunomodulation. [0003] At present, the detection of RAC1 protein is based on immunological test methods, among which the immunological detection methods include ELISA, fluoresce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/40C12N5/20G01N33/573G01N33/577
CPCC07K16/40G01N33/577G01N33/573C07K2317/56C07K2317/35G01N2333/914
Inventor 张浩洋石坚徐爱华朱国方
Owner SHAOXING SHOUREN MEDICAL HEALTH & TECH CO LTD