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Time resloved fluoroimmunoassay kit for detecting Galectin-3

An immunofluorescence, time-resolved technology, applied in the field of medical biology, can solve the problems of supply and lack of kits, and achieve the effect of short operation time, low cost and good stability

Inactive Publication Date: 2011-10-26
倪润洲 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no time-resolved immunofluorescence method specifically used to detect Galectin-3 in the world, and there is no corresponding kit provided. Therefore, the development and application of this kit is to fill the gap in this area.

Method used

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  • Time resloved fluoroimmunoassay kit for detecting Galectin-3

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Embodiment Construction

[0037] 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.

[0038] Reagent formula:

[0039] 1. Buffer:

[0040] 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;

[0041] 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;

[0042] Blocking solution: PBS containing 1% BSA, its composition is PBS, 1% BSA;

[0043] Loading buffer: PBS containing 1%BSA, its composition is PBS, 1%BSA;

[0044] Detection buffer: 50mM Tris-HCl buffer (pH 7.8), containing 1% BSA, 1% bovine globulin, 0.05% Tween 40, DTPA;

[0045] 2. Standard protein:

[0046] Commercialized r...

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Abstract

The invention discloses a time resloved fluoroimmunoassay (TRF) kit for detecting Galectin-3.The kit comprises: a coating buffer, a blocking solution, a cleaning solution, a loading buffer, a enhancement solution, a assay buffer, coated antibody (monoclonal anti-human Galectin-3 antibody), detection antibody (biotinylated anti-human Galectin-3 antibody), Eu<3+> labeled streptavidin and neomycin, a standard substance (recombinant human Galectin-3). The kit is characterized in that: the kit adopts a double antibody sandwich method and TRF principle for detecting a concentration of the Galectin-3 in a sample. The method is characterized by: coating an ELISA plate through the coated antibody; adding the sample to be detected to the ELISA plate to carry out a incubation after blocking; washing the plate, then adding the detection antibody to the plate to carry out the incubation; washing the plate again, followed by adding the Eu<3+> labeled streptavidin to carry out the incubation; washing the plate, followed by adding the enhancement solution; oscillating for 5 minutes, followed by exciting through light having a excitation wavelength of 337nm; detecting a fluorescence value of the sample at a emission wavelength of 615nm after delaying 200 microseconds; drawing a standard curve through the concentration of the standard substance and the corresponding fluorescence value, such that the concentration of the Galectin-3 in the sample to be detected can be calculated through the fluorescence value and the standard curve.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and relates to a time-resolved immunofluorescence (Time Resolved Fluoroimmunoassay, TRF) detection method, in particular to detecting the concentration of Galectin-3 protein in a sample by using the time-resolved immunofluorescence method. The increase of Galectin-3 protein concentration in human peripheral blood is closely related to the occurrence and development of pancreatic cancer, and the use of this kit can play a diagnostic role in the occurrence and development of pancreatic cancer. Background technique [0002] Galectin-3 is a β-galactoside-binding protein, a member of the lectin family. It has a CRD domain and has a high affinity with glycoproteins and glycolipids containing β-galactoside. They play a specific adhesion role and are closely related to tumor metastasis, infiltration, growth and adhesion. We use MALDI-TOF-MS mass spectrometry to find that Galectin-3 is highly express...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/64
Inventor 倪润洲倪温慨肖明兵
Owner 倪润洲
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