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Anti-SMIM15 monoclonal antibody and application thereof

A technology of SMIM15, monoclonal antibody, applied in the field of immunology

Active Publication Date: 2021-05-14
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research on protein-coding genes has gradually become an important target of basic scientific research and biomedicine, which has increased the market demand for biological reagents used in medical research and clinical diagnosis. According to the current domestic and foreign literature reports, there is no human-mouse cross-reaction. , Highly specific anti-SMIM15 monoclonal antibody can be used in the research of immunological methods

Method used

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  • Anti-SMIM15 monoclonal antibody and application thereof
  • Anti-SMIM15 monoclonal antibody and application thereof
  • Anti-SMIM15 monoclonal antibody and application thereof

Examples

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Embodiment 1

[0033] 1, the main reagent used in the following examples is:

[0034] (1) For cell fusion:

[0035] IMDM medium;

[0036] IMDM complete medium (containing 15% serum);

[0037] 2.2% methylcellulose (SIGMA; catalog number: M0262-100G);

[0038] Neonatal bovine serum; PEG1500 (Roche; catalog number: 78364);

[0039] HAT (Sigma; item number: H0262-10VL);

[0040] HT (Sigma; Cat. No.: H0137-10VL).

[0041] (2) For monoclonal cell screening:

[0042] Coating solution: sodium carbonate-sodium bicarbonate buffer (pH9.6); PBS buffer (pH7.4);

[0043] Blocking solution: PBS containing 2% milk;

[0044] Washing solution: PBS-T (0.05% Tween, PBS);

[0045]Chromogenic solution: 1% A solution + 10% B solution (A solution: 1% TMB in DMSO; B solution: 0.1% H 2 o 2 citrate buffer);

[0046] Stop solution: 2M sulfuric acid;

[0047] Secondary antibody: goat anti-mouse IgG / HRP.

[0048] (3) For the identification of monoclonal cell subtypes:

[0049] Coating antibody (Southern Bio...

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Abstract

The invention discloses an anti-SMIM15 monoclonal antibody and application thereof, belonging to the technical field of immunology. The antibody is secreted by a hybridoma cell strain 1M5. The preparation method comprises the following steps: preparing a mouse immunogen and a detection antigen; preparing the anti-SMIM15 hybridoma cell strain 1M5; screening an SMIM15 positive monoclonal antibody; carrying out subtype identification on the monoclonal antibody; detecting the specificity of the SMIM15 positive monoclonal antibody; producing a monoclonal antibody in a BALB / c mouse and purifying the monoclonal antibody; detecting the purity of the antibody by using SDS-PAGE; and subjecting the SMIM15 monoclonal antibody to functional identification and application by adopting WB, IHC and ELISA. The antibody directly secreted by the hybridoma cell strain 1M5 is immune globulin, has no toxicity, can be applied to immunoblotting and immunohistochemistry, and can also be used as an active component to prepare reagents or drugs for diagnosis and treatment of SMIM15 related diseases.

Description

technical field [0001] The invention belongs to the technical field of immunology and relates to a monoclonal antibody, in particular to an anti-SMIM15 monoclonal antibody and application thereof. Background technique [0002] Small Integral Membrane Protein 15 (Small Integral Membrane Protein 15, SMIM15, alias C5orf43) is a protein-coding gene that is commonly expressed in the colon, thyroid and other tissues of the body. It is known that SMIM15 includes 2 transcripts and 155 homologs The main function of SMIM15 is to control the synthesis of proteins under various physiological and pathological conditions, and further play a key role in the process of immune recognition and material transportation between cells. [0003] Compared with normal tissues, the expression of SMIM15 is higher in most cancer tissues, such as glioma, gastric cancer and breast cancer, but its related mechanism and research have not been reported in the literature. In recent years, as immunotherapy a...

Claims

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Application Information

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IPC IPC(8): C07K16/18C12N5/20G01N33/574G01N33/577A61K39/395A61P35/00
CPCC07K16/18G01N33/577G01N33/57407G01N33/57446G01N33/57415G01N33/57484A61P35/00C07K2317/92C07K2317/35A61K2039/505C07K2317/76C07K2317/33
Inventor 黄剑飞陆冰孙萍萍张筱静
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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