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Detection method and detection kit of early-stage irreversible damage degree of cells

A technology of damage degree and detection method, which is applied in the field of detection of early irreversible damage to cells, can solve the problems of loss of life behavior and physiological functions, cell nucleus disintegration, etc., and achieve the effect of rich content, maintenance of cell shape, and high accuracy

Active Publication Date: 2015-05-27
PROTEINT (TIANJIN) BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More importantly, from a pathological point of view, the damage of cytoskeletal proteins, cell membrane damage, and lysosome release are the three major signs of irreversible damage to cells, which eventually lead to the disintegration of the cell nucleus and the complete loss of life behavior and physiological functions

Method used

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  • Detection method and detection kit of early-stage irreversible damage degree of cells
  • Detection method and detection kit of early-stage irreversible damage degree of cells
  • Detection method and detection kit of early-stage irreversible damage degree of cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The test kit includes:

[0047] ELISA plate;

[0048] The capture antibody immobilized on the enzyme-labeled well plate, the capture antibody was purchased from Abcam Company of the United States;

[0049] Phosphate group indicator antibody, the phosphate group indicator antibody is a monoclonal antibody against the phosphorylation group of tyrosine site No. 1176 of α-fodrin; its preparation method is as follows:

[0050] 1. Synthesize short peptides with and without phosphate groups. The short peptide sequence is 11 consecutive amino acids with the 1176th amino acid as the midpoint on human α-fodrin;

[0051] 2. Inject different short peptides into different mice to induce immune responses;

[0052] 3. Fusion of mouse splenocytes with hybridoma cells;

[0053] 4. Pick monoclonal cells that recognize short peptides with phosphate groups but not short peptides without phosphate groups, that is, cells that can produce phosphate group-indicating antibodies.

[0054] α-...

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Abstract

The invention provides a detection method of damage degree of cells. In the method, chemical modification change degree of cytoskeletal protein is detected and is employed as a parameter for evaluating the damage degree during an early stage of irreversible damage of the cells, which means that a ratio value of the amount of molecules which are subjected to chemical modification change in the cytoskeletal protein to the amount of molecules which are not subjected to chemical modification change in the cytoskeletal protein is employed as a parameter for evaluating the damage degree of the cells in a to-be-detected sample. In the method, quantitative analysis of chemical modification change acting on important biological molecules, such as the cytoskeletal protein and the like, is carried out for reflecting a damage situation of the cells. The method is simple in operation, is high in sensitivity and accuracy, and has a special application value in the field of quality evaluation of organ transplantation and the like.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to a method for detecting the degree of early irreversible damage of cells. Background technique [0002] Cells are the basic structural unit of living organisms. The life activities of cells are carried out in the process of dynamic balance between internal and external environments. When cells and tissues are stimulated by intolerable harmful factors, they may cause damage. The milder cell damage is reversible. After eliminating the stimulating factor, the damaged cells can return to normal, which is usually called sublethal cell damage. Severe damage to cells is irreversible, eventually causing cell death. There are many reasons for cell damage, which can be summarized into several categories: hypoxia, chemical substances and drugs, physical factors, biological factors, nutritional imbalance, endocrine factors, immune response, genetic variation, aging and so on. [0003] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6842
Inventor 李捷
Owner PROTEINT (TIANJIN) BIOTECHNOLOGY CO LTD
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