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Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549

A lung adenocarcinoma cell and fluorescent antibody technology is applied in the field of preparing rabbit anti-human lung adenocarcinoma cell A549 fluorescent antibody to achieve the effect of improving sensitivity and preventing antibody activity from decreasing or degeneration

Inactive Publication Date: 2015-01-28
ZUNYI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There is no report about the production of A549 monoclonal antibody. The A549 antibody used in the present invention is the polyclonal IgG extracted and purified by our laboratory after immunizing rabbits with A549 cells.

Method used

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  • Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549

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Embodiment Construction

[0023] The specific embodiment of the present invention, its technical scheme and steps are stated as follows again now:

[0024] 1. Culture and subculture of A549 cells: cell culture at 37°C in 5% CO 2 In the incubator, observe the growth of A549 cells under an inverted microscope

[0025]When the cell growth density was close to 80%, the cells were digested and passaged. Pour out the old culture medium in the bottle, add 1ml of trypsin, shake the bottle slightly and pour it out quickly, then add 1-2ml of trypsin, observe under the inverted microscope after 1-3min when the cells become round and start to fall off, add 4 -5ml of serum-containing medium to stop the digestion, blow the cells gently with a pipette to the cell suspension, pass passage at a ratio of 1:3, then add 1640 culture medium containing 10% FBS to 5ml, and stand at 37°C, 5%CO 2 cultured in an incubator.

[0026] 2. Preparation of A549 antigen: After digesting the adherent A549 cells with trypsin, a single...

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Abstract

The invention provides preparation of a fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549. Immunogen of human lung adenocarcinoma cells A549 is injected into a rabbit to prepare a polyclonal antibody of the rabbit anti-human lung adenocarcinoma cells A549. according to the invention, the adherent A549 cells are digested by pancreatin to prepare single-cell suspension, the cell density is adjusted, formaldehyde is added for inactivate, the A549 cell immunogen is subcutaneously injected in the rabbit (0.5 ml) at multipoint at the first, third, seventh and fourteenth days, and blood is extracted from the heart after immune injection of 30 days to separate the serum. IgG is roughly extracted by ammonium sulfate salt fractionation, IgG is purified by dextrangel, and the fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 is prepared from a dialysis method. The antibody can be used for identifying multiple epitopes of the same antigen, improving the detection sensitivity under the same condition, can be used for verifying human lung adenocarcinoma cells and diagnosing immunopathogenesis of human lung adenocarcinoma, and has good social benefits and market prospects.

Description

technical field [0001] The invention relates to the technical field of medicine and biology, in particular to a preparation of rabbit anti-human lung adenocarcinoma A549 fluorescent antibody. Background technique [0002] Primary lung cancer is one of the most common malignant tumors in the world, and its case fatality rate ranks first in the cause of cancer death in the world; its causes include smoking, air pollution, certain occupational factors, genetic factors, etc., ranking third in the country In the second cause of death survey, lung cancer accounted for 22.7% of the national cancer death causes, ranking first among the national cancer death causes. Lung cancer can be roughly divided into two types: non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). Among them, non-small cell lung cancer accounts for about 80%-85% of the total number of lung cancers. Compared with small cell lung cancer, the growth of its cancer cells Division is slower and diffus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30C07K16/06C07K1/36C07K1/30C07K1/16C07K1/13
Inventor 孙万邦王少慧郭锦锦宋明英
Owner ZUNYI MEDICAL UNIVERSITY