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Galectin-3 detection nanometer magnetic bead sorting-time resolved immunofluorescence kit

A time-resolved, immunofluorescence technology, applied in biological testing, measurement devices, analytical materials, etc., can solve the problems of no nano-magnetic bead sorting-time-resolved immunofluorescence kit, etc., and achieve low cost, good stability, and easy operation short time effect

Inactive Publication Date: 2015-04-15
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] At present, there is no relevant report in the world, and there is no nano-magnetic bead sorting-time-resolved immunofluorescence kit specifically for the detection of Galectin-3. Therefore, the development and application of this kit fills the gap in this regard.

Method used

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  • Galectin-3 detection nanometer magnetic bead sorting-time resolved immunofluorescence kit
  • Galectin-3 detection nanometer magnetic bead sorting-time resolved immunofluorescence kit
  • Galectin-3 detection nanometer magnetic bead sorting-time resolved immunofluorescence kit

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Embodiment Construction

[0050] Some terms and abbreviations in the present invention: TRF: time-resolved immunofluorescence; BSA: bovine serum albumin; Tween-20: Tween 20; Tween-40: Tween 40; PBS: phosphate buffered saline; TBS: Tris- HCl buffer; bovine globulin: bovine globulin; DTPA: diethylenetriaminepentaacetic acid.

[0051] Reagent formula:

[0052] 1. Buffer:

[0053] Coating buffer: phosphate buffered saline (PBS), its composition is 140mM NaCl, 2.7mM KCl, 10mM NaCl 2 HPO 4 , 1.8mM KH 2 PO 4 , the pH value is 7.4;

[0054] Washing solution: Tris-HCl buffer (TBS) containing 0.05% Tween-20, its composition is 50mM Tris-HCl buffer (pH 7.8), 0.05% Tween-20;

[0055] Blocking solution: PBS containing 1% BSA, its composition is PBS, 1% BSA;

[0056] Loading buffer: PBS containing 1%BSA, its composition is PBS, 1%BSA;

[0057] Detection buffer: 50mM Tris-HCl buffer (pH 7.8), containing 1% BSA, 1% bovine globulin, 0.05% Tween 40, DTPA;

[0058] 2. Standard protein:

[0059] Commercialized r...

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Abstract

The invention relates to a Galectin-3 detection nanometer magnetic bead sorting-time resolved immunofluorescence kit, which comprises superparamagnetic composite micro-particles with a particle size of 30 nm, a coupling liquid, a blocking solution, a washing liquid, a loading buffer, a fluorescence enhancement solution, a detection buffer, a capture antibody, a detection antibody, Eu<3+> labeled streptavidin and a standard substance. The use of the kit comprises: adopting nanometer magnetic bead sorting and time resolved immunofluorescence adopting a double antibody sandwich method as the base to detect the concentration of Galectin-3 protein in a sample, adopting nanometer magnetic beads coupled with the Galectin-3 capture antibody to enrich the Galectin-3 in the sample, adding the detection antibody, incubating, washing, adding the Eu<3+> labeled streptavidin, incubating, washing again, adding the fluorescence enhancement solution, vibrating for 5 min, adopting light with a wavelength of 337 nm to excite, delaying for 200 microseconds, detecting the emitted fluorescence value at a wavelength of 615 nm, and obtaining the concentration of the Galectin-3 in the sample to be detected through the fluorescence value and the corresponding standard curve, wherein the sample to be detected can further be used for detecting the next target protein.

Description

[0001] technical field [0002] The invention belongs to the field of medical biotechnology, and relates to a tandem detection method of nano-magnetic bead sorting and time-resolved immunofluorescence, specifically using nano-magnetic beads coupled with Galectin-3 antibody to specifically capture and detect Galectin-3 protein in a sample , and then time-resolved immunofluorescence method was used to further detect the concentration of captured Galectin-3 protein. The increase of Galectin-3 protein concentration in human peripheral blood is closely related to the occurrence and development of pancreatic cancer. The use of this kit can play a diagnostic role in the occurrence and development of pancreatic cancer, realize the repeated use of serum samples, and improve the detection of serum samples. Detection sensitivity of Galectin-3. [0003] Background technique [0004] Galectin-3 is a galectin expressed in the cytoplasm, cell membrane and mitochondria, and can be secrete...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/54326G01N21/6408G01N21/6486G01N33/54346G01N33/57438G01N33/68
Inventor 倪润洲
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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