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A method for quantitative detection of protein concentration by flow cytometry

A technology of protein concentration and flow cytometry, which is applied in the field of quantitative detection of protein concentration by flow cytometry, can solve the problems of cumbersome operation steps, and achieve the effect of convenient and fast operation, high specificity and sensitivity, and accurate quantification

Active Publication Date: 2019-05-14
王博
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Problems solved by technology

This method has the advantages of being fast, sensitive, simple and the carrier is easy to standardize, but the specific operation steps are cumbersome, and each reaction system can only detect one indicator, and the lower limit of detection of this method is at the ng / ml level

Method used

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  • A method for quantitative detection of protein concentration by flow cytometry
  • A method for quantitative detection of protein concentration by flow cytometry
  • A method for quantitative detection of protein concentration by flow cytometry

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Embodiment Construction

[0023] In order to better understand and implement the present invention, the present invention will be further described in detail below with reference to specific implementation examples.

[0024] A method for quantitatively detecting protein concentration by flow cytometry, combining fluorescence immunomagnetic microspheres and flow cytometry to detect specific protein concentration, selecting rhodamine 123 labeled magnetic microspheres and PE-cy7 labeled detection antibody, the specific method is as follows:

[0025] 1. Preparation of fluorescent magnetic microspheres: fluorescent dyes are modified on magnetic microspheres to prepare fluorescent magnetic microspheres. The specific preparation method is as follows:

[0026] (1) Weigh 3 mg of rhodamine 123 (Rh123) dye and dissolve it in 10 mL of deionized water to prepare a 1 mmol / L dye mother liquor. Here rhodamine 123 or PE-cy7 is an example. This method can select any two fluorescent dyes of different channels of the flow cytome...

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Abstract

The invention discloses a method for quantitatively detecting protein concentration by a flow cytometer, and specific protein concentration is detected by the combination of fluorescence immunological magnetic microspheres and flow cytometric detection. The method comprises the specific steps: preparing a fluorescence detection antibody; preparing fluorescence immunological magnetic microspheres and calibrating magnetic microspheres; making a standard article working curve, and acquiring actual protein concentration through unknown sample average fluorescence strength. The flow cytometry and the fluorescence immunological magnetic microspheres are combined for detecting protein concentration; specific protein molecular concentration is detected by using the method, a reaction system is small, measuring is more accurate, a detection lower limit is pg / ml level, specificity and sensitivity are high, operating is more convenient and quick, operating steps are simpler, incubation is direct without stepwise incubation and washing so that reaction time is shortened, detection results are obtained within a shorter time, and quantitating is more accurate.

Description

Technical field [0001] The invention relates to a method for quantitative detection of protein concentration, in particular to a method for quantitative detection of protein concentration by a flow cytometer. Background technique [0002] Biological macromolecules are multi-molecular systems formed by polymerization of organic compounds with simple structures and relatively low molecular weights, including proteins, nucleic acids, lipids and carbohydrates. Biological macromolecules have various biological activities and play an important role in biological metabolism, so they are the basic materials of life. In particular, protein, which is the basic functional unit of human tissues, is the main undertaker of life activities, and at the same time exerts biological functions through complex structural characteristics, such as providing energy and substances needed for life, promoting growth and development, and generating immune functions. [0003] The research on protein requires ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14G01N33/68
CPCG01N15/1468G01N33/68
Inventor 王博
Owner 王博
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