Kit used for detecting TPD52L1 protein, and preparation method thereof

A kit and protein technology, applied in the field of chemiluminescent immunoassay, can solve the problems that cannot satisfy the ultra-trace detection of TPD52L1 protein, and achieve the effect of improving the luminescent signal

Active Publication Date: 2014-02-12
SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0022] The present invention overcomes the technical problem that the current chemiluminescence method cannot meet the ultra-trace detection of TPD52L1 protein, that is, combines the chemiluminescence technology with the biotin-avidin immunomagnification technology and the water-soluble 5,10,15-tris(4-pyridyl )-20-R-carboxyporphyrin is used as a marker to construct a chemiluminescent immunoassay to detect TPD52L1 protein with high sensitivity, and a kit for detecting TPD52L1 protein and its preparation method are provided

Method used

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  • Kit used for detecting TPD52L1 protein, and preparation method thereof
  • Kit used for detecting TPD52L1 protein, and preparation method thereof
  • Kit used for detecting TPD52L1 protein, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Preparation of the kit for detecting TPD52L1 protein of the present invention

[0057] 1. Preparation of calibrator matrix solution

[0058]

[0059] Weigh the above reagents into a clean container, add double distilled water to make up the volume, dissolve and mix well, and measure the pH value. Store at 2-8°C after labeling.

[0060] 2. Preparation of TPD52L1 protein calibrator

[0061] Prepare calibrator with calibrator matrix solution and pure TPD52L1 protein, the concentrations are 0, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100ng / mL, (wherein 0ng / mL is calibrator matrix solution, excluding TPD52L1 protein), prepare a series of concentration calibration products, each concentration calibration product is divided into 1 mL / bottle with 3 mL size vials, freeze-dried, and stored at -20 °C.

[0062] 3. Preparation of formula diluent

[0063]

[0064] 4. Preparation of streptavidin-coated solid-phase magnetic particles

[0065] (1) Weigh 250 mg of amino-t...

Embodiment 2

[0095] Example 2 Preparation of the kit for detecting TPD52L1 protein of the present invention

[0096] 4. Preparation of streptavidin-coated solid-phase plastic particles

[0097] (1) Weigh 250 mg of amino-terminal plastic particles and place them in a beaker;

[0098] (2) Add 5 mL each of methanol and acetone to wash repeatedly 5 times;

[0099] (3) centrifugal separation, discard the supernatant;

[0100] (4) Add 10 mL of 0.2% 1,4-phenylene diisothiocyanate (PDITC) solution dissolved in DMF, place in a constant temperature oscillator, and react for 10 h;

[0101] (5) Wash the reacted plastic particles 6 times repeatedly with acetone and ultrapure water;

[0102] (6) Take the newly prepared plastic particles activated by PDITC and place them in a 30mL centrifuge tube;

[0103] (7) Add 25 mL of PBS, add 4 mg of streptavidin, and place in a constant temperature shaker to react for 20 min;

[0104] (8) Centrifuge, measure the OD value of the solution at 280nm before and af...

Embodiment 3～6

[0106] Examples 3-6 Prepare the kit for detecting TPD52L1 protein of the present invention

[0107] Monoclonal antibodies were replaced with polyclonal antibodies, genetically engineered antibodies, proteins, macromolecular organic compounds, and the rest were prepared in the same way as in Example 1 to prepare the TPD52L1 protein detection kit of the present invention.

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Abstract

The invention discloses a kit used for detecting TPD52L1 protein, and a preparation method thereof, and belongs to the field of chemiluminescence immune assay technology. The kit is capable of solving technical problems of existing TPD52L1 protein ultramicro detection technology. The kit comprises: 1) a TPD52L1 protein calibration material; 2) a streptavidin-coupled solid phase carrier; 3) a biotinylated TPD52L1 protein ligand; 4) a 5, 10, 15- tris(4-pyridyl)-20-R-carboxylporphyrin coupling labeled TPD52L1 protein ligand; and 5) a chemiluminescence substrate of 5, 10, 15-tris(4-pyridyl)-20-R-carboxylporphyrin.

Description

technical field [0001] The invention relates to the technical field of chemiluminescence immunoassay, in particular to a kit for detecting TPD52L1 protein and a preparation method thereof, which combines biotin-avidin immunomagnification technology and chemiluminescence immunoassay technology. Background technique [0002] TPD52L1 protein is a protein encoded and expressed by the TPD52L1 gene, which is a member of the D52 series gene family and expressed in human breast cancer cells. The encoded protein contains a coiled-coil domain. Functions in cell proliferation and calcium signaling protein. It also interacts with mitogen-activated protein kinase kinase 5 (MAP3K5 / ASK1) and actively regulates MAP3K5, inducing apoptosis. In the process of scientific research and production, it is necessary to measure the change of TPD52L1 protein content in the solution to monitor the experimental conditions and monitor the production process. [0003] At present, TPD52L1 protein assay ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/532G01N33/54326G01N33/544G01N33/6803
Inventor 陈秀发李勇徐海伟胡庆锋常立峻
Owner SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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