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Quantum dot mark based immune blotting detection method

A technology of immunoblotting and detection method, which is applied in the field of protein detection, can solve the problems of complex design platform, low fluorescence stability, and low detection sensitivity, and achieve the effects of saving time and cost, increasing photostability, and improving sensitivity

Inactive Publication Date: 2008-08-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

Low fluorescence stability makes it difficult to image over long periods of time, resulting in low detection sensitivity
Because different chromophores require different wavelength excitation sources and require a very complex design platform, multi-channel technology is also not possible
In addition, the dye's broad emission spectrum allows multiple pathways to overlap and interfere with each other

Method used

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Embodiment Construction

[0024] The embodiments of the present invention are described in detail below: the present embodiment is implemented under the premise of the technical solution of the present invention, and detailed implementation and specific operation process are provided, but the protection scope of the present invention is not limited to the following implementation example.

[0025] The expression of BRCAA1 protein was detected by immunoblotting detection method labeled with quantum dots.

[0026] Quantum dots with carboxyl groups on the surface (emission wavelengths of 520nm (green), 600nm (orange), 620nm (red orange) and 680nm (red)) were dispersed in a borate buffer solution (ph=9.18), and disulfide was added Imine (EDC) and goat anti-rabbit secondary antibody were mixed by vortexing and oscillated evenly, and reacted at room temperature for 3 hours. The reaction solution was centrifuged at high speed and washed several times with phosphate buffer solution (PBS, 0.1M, pH=7.8) to obta...

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Abstract

The present invention relates to a method for detecting immunoblotting based on quantum dot labeling. Firstly, connecting quantum dot with carboxyl or amido with second antibody by chemical reaction to form compound with quantum dot label; employing polyacrylamide gel electrophoresis, analytes is protein, probe is antibody, quantum dot labeled second antibody is used to show color. Protein sample separated by PAGE is transferred to solid carrier, the protein or polypeptide on solid carrier as antigen is processed immunoreaction with corresponding antibody, and then reacted with quantum dot labeled second antibody, under the UV lamp, quantum dot emitting fluorescence to detect electrophoresis separated specific objective gene expression protein component. Immunoblotting image is gained by filter equipped of simple UV imaging system. The present invention employs quantum dot connected with second antibody as excellent fluorescence imaging agent of immunoblotting to enhance the sensitivity and precision.

Description

technical field [0001] The invention relates to a protein detection method in the field of nanotechnology, in particular to a quantum dot-based immunoblotting detection method. Background technique [0002] Conventional western blot (Western Blots) uses chemiluminescence and radiation techniques. Western blot analysis techniques using traditional organic fluorescent chromophores have inherent limitations. Low fluorescence stability makes it difficult to image over long periods of time, resulting in low detection sensitivity. Multi-channel technology is also not possible because different chromophores require different wavelengths of excitation light sources and a very complex design platform is required. In addition, the broad emission spectrum of dyes allows multiple pathways to overlap and interfere with each other. [0003] Quantum dots (QDs) specifically refer to semiconductor nanoparticles whose radius is smaller than or close to the exciton Bohr radius. Semiconducto...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/561G01N21/64
Inventor 贺蓉崔大祥高峰
Owner SHANGHAI JIAO TONG UNIV
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