Immunization grouping analysis and detection method based on quantum point

A technology of immunohistochemistry and detection methods, which is applied in the direction of analyzing, analyzing materials, and measuring devices by causing chemical reactions to occur in materials, and can solve problems such as restrictions on popularization and application, fading fluorescence intensity, and difficulty in long-term storage of results. Increased stability, improved sensitivity, time and cost savings

Inactive Publication Date: 2009-10-07
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

However, currently commonly used fluoresceins are mostly organic dyes, and there are disadvantages such as the fluorescence intensity gradually fades with time, and the results are not easy to store for a long time, which is limited in popularization and application.

Method used

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Embodiment

[0025] The detection method is to analyze the expression of BRCAA1 protein in various tumor tissues. The primary antibody is a rabbit IgG polyclonal antibody against the neoantigen epitope of human BRCAA1, and the secondary antibody is a goat anti-rabbit IgG monoclonal antibody labeled with quantum dots. Solution (PBS) was used as a negative control instead of the primary antibody.

[0026] The formula of borate buffer solution is:

[0027] Sodium tetraborate decahydrate 3.81g, add double-distilled water to make the volume to 1000mL, pH is 9.18.

[0028] The formula of phosphate buffer solution is:

[0029] Sodium chloride 8g

[0030] Potassium chloride 0.2g

[0031] Disodium hydrogen phosphate 1.44g

[0032] Potassium dihydrogen phosphate 0.24g

[0033] Add double distilled water to 1000ml, and adjust the pH to 7.4.

[0034] Step 1, disperse the water-soluble CdTe quantum dots with carboxyl groups on the surface in borate buffer solution (pH=9.18), add diimide sulfide (...

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Abstract

An immunization grouping analysis and detection method based on quantum point belongs to the nano-material field, comprises the following steps: (1) dispersing quantum points into borate buffer solution, adding sulfurated diimine and second antibody, mixing, reacting, centrifugating, washing, and obtaining second antibody compound with quantum point marks; wherein, the mole ratio of the quantum points, the sulfurated diimine and the second antibody is: 60: 15: 1; or (2) dispersing the quantum points into phosphoric acid buffer solution containing 1.25 % of glutaraldehyde by volume fraction, dispersing, reacting, adding second antibody, reacting, centrifugating, washing, and obtaining second antibody compound with quantum point marks; wherein, the mole ratio of the quantum points, the glutaraldehyde and the second antibody is: 60: 15: 1; then performing immunization reaction and observation according to the standard immunization grouping detection method. The method in the invention performs observation and judgement directly by fluorescence microscope, is simple and easy, can increase the fluorescence stability, and improve analysis accuracy and detection sensitivity.

Description

technical field [0001] The invention relates to a detection method in the field of nanotechnology, in particular to an immunohistochemical analysis detection method based on quantum dots. Background technique [0002] Immunohistochemistry is the application of the basic principle of immunology - antigen-antibody reaction, that is, the principle of specific binding of antigen and antibody, which is determined by the color development of the chromogenic agent (fluorescein, enzyme, metal ion, isotope) of the labeled antibody through a chemical reaction. Tissue intracellular antigens (polypeptides and proteins) for localization, qualitative and quantitative research. Since the immune complex formed after the combination of antibody and antigen is colorless, it is also necessary to display the antigen-antibody reaction site by means of histochemical methods. Through the antigen-antibody reaction and color reaction, the chemical components in the cells or tissues can be displayed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/536G01N21/76
Inventor 贺蓉崔大祥高峰徐昕
Owner SHANGHAI JIAO TONG UNIV
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