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ELISA kit for quantitatively detecting tight junction related protein Occludin

A connection-related and quantitative detection technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of expensive detection and discomfort, and achieve easy operation, good detection accuracy and precision, and reduce costs Effect

Pending Publication Date: 2020-02-11
安徽恩禾生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Because enhanced CT needs to inject contrast agent when using it, a very small number of people may have allergic reactions to contrast agent. According to the general report of patients, there will be obvious discomfort after injection of contrast agent; and the detection cost is expensive; Upgraded on the basis of CT, but also has the general disadvantages of CT

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  • ELISA kit for quantitatively detecting tight junction related protein Occludin
  • ELISA kit for quantitatively detecting tight junction related protein Occludin

Examples

Experimental program
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Embodiment 1

[0034] This embodiment provides the composition and preparation method of an ELISA kit for the quantitative detection of the tight junction-related protein Occludin:

[0035] 1. The composition of the kit

[0036] 1) Occludin-coated plate: 48-well / 96-well microplate coated with Occludin monoclonal antibody

[0037] 2) Occludin calibrator 1 set A-F: derived from human serum, concentration 120±12ng / L, 80±8ng / L, 40±4ng / L, 20±2ng / L, 10±1ng / L, 0ng / L, available Traceable to the measurement procedure selected by the manufacturer. 1×0.5ml

[0038] 3) Occludin-binding antibody: rabbit anti-human Occludin polyclonal antibody. 1×2.5ml / 1×5.0ml

[0039] 4) Occludin enzyme conjugate: HRP-labeled goat anti-rabbit IgG. 1×5.0ml / 2×5.0ml

[0040] 5) Occludin quality control substance: derived from human serum, the quality control range is 90ng / L±13.5ng / L. 1×0.5ml

[0041] 6) Color developer A: the main component is urea peroxide. 1×5.0ml

[0042] 7) Chromogenic agent B: the main ingredie...

Embodiment 2

[0085] This embodiment provides a detection method of an ELISA kit for the quantitative detection of the tight junction-related protein Occludin comprising the following steps:

[0086] 1) Preparation of washing solution: add 20ml of concentrated washing solution into a 500ml volumetric flask, then add 480ml of distilled water, shake to fully dissolve the concentrated washing solution;

[0087] 2) After the kit is equilibrated to room temperature, take out the required microwell strips and fix them on the plate rack, and compile the sequence;

[0088] 3) Adding samples: first add 50ul of the sample to be tested / Occludin calibrator / Occludin quality control substance, then add 50ul of Occludin-binding antibody to the corresponding well, pat to mix, and set a blank well;

[0089] 4) Incubation: seal the strips with parafilm, and incubate at 37°C for 30 minutes;

[0090] 5) Plate washing: take out the reaction plate, shake off all the liquid in the plate, fill each well with wash...

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Abstract

The invention relates to the technical field of detection kits, and particularly discloses an ELISA kit for quantitatively detecting tight junction related protein Occludin. The kit comprises an elisaplate coated with an Occludin coated antibody, HRP labeled goat anti-rabbit IgG, a rabbit anti-human Occludin polyclonal antibody, a washing solution, a sample diluent, a color developing agent, a stop solution, a quality control material and a standard substance, the kit is based on the Occludin coated antibody and the goat anti-rabbit IgG, and an ELISA method for Occludin protein detection is established by adopting a double antibody sandwiching method. The kit overcomes the defects in the prior art, can be used for effectively detecting the level of Occludin protein in a serum sample, is simple, convenient and rapid to operate, has good detection accuracy and precision, high specificity, good sensitivity and good stability, and can be used for rapidly and timely helping to diagnose theillness state and monitor prognosis.

Description

technical field [0001] The invention relates to the technical field of detection kits, in particular to an ELISA kit for quantitatively detecting tight junction-related protein Occludin. Background technique [0002] The blood-brain barrier (BBB) ​​is an important structure to maintain the stability of the unique internal environment of the central nervous system, and it is the premise and basis for the neurons of the central nervous system to carry out various physiological activities. The basic structure of the blood-brain barrier includes: brain microvascular endothelial cells and their tight junctions (TJ), endothelial basement membrane and astrocyte terminal feet. Among them, brain microvascular endothelial cells and their tight junctions are the core structure of the blood-brain barrier. [0003] The basic ultrastructural characteristics of tight junctions in various tissues and organs are similar, and they are all manifested as: under the transmission electron micros...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/543
CPCG01N33/6893G01N33/581G01N33/577G01N33/543G01N2333/705G01N2800/2871
Inventor 余旭亮张儒强
Owner 安徽恩禾生物技术有限公司
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