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ELISA kit for detecting pygo2 protein content in human serum, method of use and application

A protein and reagent technology, applied in the fields of biotechnology and medicine, to achieve high sensitivity and high accuracy

Inactive Publication Date: 2019-01-29
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, there is no kit for detecting serum levels of Pygo2 protein by ELISA

Method used

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  • ELISA kit for detecting pygo2 protein content in human serum, method of use and application

Examples

Experimental program
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Effect test

Embodiment 1

[0035] [Example 1] Detection of Pygo2 protein in serum of patients with colorectal cancer using ELISA kit

[0036] 1. Sample processing: collect blood from the patient, put it in a centrifuge tube, centrifuge at 2500rpm for 10min after the blood coagulates, obtain the serum sample to be tested, and store it at -20°C to avoid repeated freezing and thawing.

[0037] 2. Take the coated ELISA plate and wash the ELISA reaction plate 5 times with PBST; dilute the negative and positive control sera and the serum to be tested 1:100 times, add 100 μL to the reaction well of the ELISA plate, and repeat the well for each group of samples After incubating at 37°C for 1 hour, wash each reaction well of the ELISA plate 5 times with PBST; dilute the horseradish peroxidase-labeled anti-Pygo2 protein antibody in the blocking solution, add it to the plate, and incubate at 37°C. Use PBST to wash each reaction well of the ELISA microplate plate 5 times; add 50 μL each of substrate solutions A and...

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Abstract

The invention discloses an ELISA kit for detecting content of Pygo2 protein in human serum as well as a using method and application thereof. A double-antibody sandwich method ELISA kit based on the Pygo2 protein is prepared from the following raw materials: an ELISA Plate coated with antibodies with resistance to the Pygo2 protein, Pygo2 standard antigens, negative and positive control fluid, enzyme-labeled antibodies, a PBST buffer solution, a sealed buffer solution, a substrate solution A, a substrate solution B and a stop solution. The enzyme linked immunosorbent assay (ELISA) kit for detecting the content of the Pygo2 in the human serum based on the double-antibody sandwich method is capable of accurately and quantitatively detecting the content of the Pygo2 in the human serum; the detection method can be used as a method for simply and rapidly diagnosing and monitoring disease course development of the diseases such as colorectal cancer, brain glioma and the like without trauma.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and relates to an ELISA kit for detecting the Pygo2 protein content in human serum, a use method and application. Background technique [0002] The Wnt signaling pathway widely exists in invertebrates and vertebrates, and is a highly conserved signaling pathway in the evolution of species. Wnt signaling plays a vital role in the early development of animal embryos, organ formation, tissue regeneration and other physiological processes. If the key protein in this signaling pathway is mutated or abnormally expressed, leading to abnormal activation of the signal, it may induce cancer. The Wnt signaling pathway includes the classic Wnt signaling pathway and the non-canonical Wnt signaling pathway. In the classic pathway, the Wnt-β-catenin signaling pathway, the Wnt factor inhibits intracellular free cells by activating the Frizzle / LRP5 / 6 synergistic receptors on the cell membrane. The pho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/574G01N33/543
CPCG01N33/54306G01N33/57407G01N33/57419G01N33/57484G01N33/68G01N2333/47
Inventor 秦文英唐景峰周策凡张毅李舜尧何文早陈兴珍胡婷
Owner HUBEI UNIV OF TECH