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Kit for detecting NK cell activity

A NK cell and active technology, applied in the biological field, can solve the problems of high natural release rate, sensitivity and specificity of influence method, strong fluorescence background, etc., and achieve the effect of saving detection cost and detection time.

Active Publication Date: 2014-11-05
北京海思特医学检验实验室有限公司
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Problems solved by technology

The disadvantage is that the content of alkaline phosphatase in the target cells is low or some undead cells can release it by itself, which affects the sensitivity and specificity of the method
In addition, the lactate dehydrogenase molecule is relatively large, and it is only released when the target cell membrane is completely destroyed, so it cannot reflect the effector function earlier
Fluorescence analysis often encounters high natural release rate of cells and strong fluorescence background, which affects sensitivity
The operation of radioimmunoassay is simple, fast and quantitative, but the disadvantage is that the natural release rate is high, the number of target cells required is large, and the half-life is short

Method used

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  • Kit for detecting NK cell activity
  • Kit for detecting NK cell activity

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Embodiment Construction

[0016] The specific implementation of the present invention will be further described below in conjunction with the examples, and the advantages and characteristics of the present invention will become clearer along with the description. However, these embodiments are only exemplary and do not constitute any limitation to the scope of the present invention.

[0017] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0018] The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0019] NK cells contain a large number of enzymes. When the body encounters target cells, NK cells can quickly recognize, synthesize and secrete a large amount of cytokines, enzymes, perforin and other active substances to lyse and kill target cells. Based on the above theory, the present invention sets up a combined detection method of EGFP-K562 / PBMNC / 7-AAD-percp-cy5.5, 7...

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Abstract

The invention provides a kit for detecting NK (natural killer) cell activity. The kit includes a PBS buffer solution, a lymphocyte separation medium, a RPMI1640 culture solution, a fluorescent dye EGFP, an antibody 7-AAD-percp-cy5.5, and a fluorescent dye's carrier pEGFP-N1 plasmid. The invention also provides a method for detection of NK cells by the kit. The method provided by the invention can save detection time and detection cost, and has high sensitivity.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a kit for detecting NK cell activity. Background technique [0002] Natural killer cells (NK) are important immune cells in the body, not only related to anti-tumor, anti-viral infection and immune regulation, but also involved in the occurrence of hypersensitivity and autoimmune diseases in some cases. Maturation depends on the bone marrow and thymus microenvironment. The function of NK cells is mainly realized in the following ways. NK cell natural killing activity: NK cells recognize target cells (some tumor cells, virus-infected cells, some self-organized cells, parasites, etc.), then NK cells release perforin, cytotoxic factors, and activated NK cells release TNF factors , so as to achieve the possibility of lysing and killing target cells. NK cell cytotoxicity (ADCC effect), mainly due to the FcγRⅢA on the surface of NK cells, which mainly binds to the Fc segments of human IgG1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/64
CPCG01N15/14G01N21/6402G01N33/53G01N2800/00
Inventor 张娟王显凤刘丽媛吴纯斌秦晓明王绪华梁超陈忠方国伟黄士昂
Owner 北京海思特医学检验实验室有限公司
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