Magnetic bead immunodetection kit for human colorectal cancer tumor stem cell markers CD44 and CD133

A technology for tumor stem cells and colorectal cancer, applied in the field of magnetic bead immunoassay kits, can solve the problems of high price and cumbersome detection process, and achieve the effect of low cost, high price and short detection time

Inactive Publication Date: 2016-10-26
JINLIN MEDICAL COLLEGE
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Problems solved by technology

[0007] The present invention aims to solve the problems of the existing detection kits for colorectal cancer, such as high price and cumbersome detection process, and proposes a magnetic bead immunoassay kit for human colorectal cancer tumor stem cell markers CD44 and CD133

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  • Magnetic bead immunodetection kit for human colorectal cancer tumor stem cell markers CD44 and CD133
  • Magnetic bead immunodetection kit for human colorectal cancer tumor stem cell markers CD44 and CD133
  • Magnetic bead immunodetection kit for human colorectal cancer tumor stem cell markers CD44 and CD133

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Embodiment Construction

[0033] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0034] 1. CD133 gene cloning and construction of expression cells

[0035] Primers were designed according to the CD133 gene sequence, and HindⅢ and BamHI restriction sites were introduced into the cDNA of CD133 molecule by PCR. '-TAAGGATCCCATATGTTGTGTGATGGGCT-3' (with Bam HI restriction site). The PCR reaction conditions were pre-denaturation at 94°C for 5 min, denaturation at 94°C for 45 s, annealing at 58°C for 45 s, extension at 72°C for 45 s, and 35 cycles; then extension at 72°C for 10 min. The PCR amplification product was recovered, the PCR product and the pEGZ-Term vector plasmid were double-digested with restriction endonucleases Bam HI and Hind III, the purified product was ligated with T4 ligase, and the competent E. coli Top10 was transformed, and the plasmid was extracted after shaking overnight. Validation by double enzyme digestion (se...

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Abstract

The invention discloses a magnetic bead immunodetection kit for human colorectal cancer tumor stem cell markers CD44 and CD133, and belongs to the field of immunoglobulin. The kit comprises a Solution I, protease K, a fluorescent CD44 antibody and a magnetic bead CD133 antibody. Technology combining magnetic beads and fluorescence radiation is adopted in the kit for detecting CD44 and CD133 on cytomembranes, thereby saving tedious operation of a flow cytometry. A small quantity of a pathological sample of a patient is added in a kit with the magnetic bead antibody and the fluorescent antibody well prepared and is detected through a fluorescence microscope to get a result. The kit is simple to use, short in detection time and low in cost and has bright application prospects.

Description

technical field [0001] The invention belongs to the field of immunoglobulins, in particular to a magnetic bead immunoassay kit for human colorectal cancer tumor stem cell markers CD44 and CD133. Background technique [0002] Magnetic bead separation technology: Magnetic bead is a kind of carrier microsphere with a certain magnetic and special structure, which is formed by combining magnetic inorganic particles and organic polymers by a certain method. Magnetic bead separation technology is a molecular biological separation technology, which utilizes the surface-modified magnetic particles to bind biomolecules or cells for separation, and can efficiently enrich the target to be separated or detected. The method is convenient, fast, with high recovery rate and strong selectivity. [0003] Fluorescent antibody technology: Fluorescent antibody technology is a technology for antigen localization by fluorescently labeled antibodies. Compared with other serological methods for id...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/574
CPCG01N33/577G01N33/57419G01N33/57446G01N33/57484G01N2333/70503G01N2333/70585
Inventor 黄晓东沈楠齐玲王艳春张洪嘉吴畏难
Owner JINLIN MEDICAL COLLEGE
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