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Proteome sample processing kit box and processing method, and application

A proteome and processing method technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of cumbersome and long-time proteome sample processing, and achieve the effect of saving time and simplifying the processing process.

Inactive Publication Date: 2019-09-06
北京谷海天目生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to provide a proteome sample processing kit, processing method and application to solve the technical problems of cumbersome proteome sample processing procedures and long time in the prior art

Method used

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  • Proteome sample processing kit box and processing method, and application
  • Proteome sample processing kit box and processing method, and application
  • Proteome sample processing kit box and processing method, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The kit of this embodiment contains four solutions A, B, C, and D, and the ingredients are as follows in Table 1:

[0042] Table 1

[0043] Solution name Element A Tris-HCl pH=8.8 B 400mM CAA solution C 100mM TCEP solution D 10% (W / V) DOC solution

[0044] Before proteome sample processing, the three solutions of A, B, and C in the kit and the mass spectrometer water were prepared according to the formula of 1:1:1:6 to make a mixed solution, and finally 1 part of D solution was added, mixed evenly, and prepared Complete treatment solution.

[0045] Samples of this example: gastric cancer tissue and adjacent normal tissue

[0046] The operation is as follows:

[0047] 1. Weigh the empty EP tube and mark it. Each pair of cancer tissue (Tumor, T) and paired adjacent normal tissue (Nearby tissue, N) samples were guaranteed to be processed in parallel at the same time. After the tissue samples were taken out from -80°C, they were ...

Embodiment 2

[0056] The kit of this embodiment contains four solutions A, B, C, and D, and the ingredients are as follows in Table 2:

[0057] Table 2

[0058] Solution name Element A Tris-HCl pH=8.8 B 400mM CAA solution C 100mM TCEP solution D 10% (W / V) DOC solution

[0059] Before proteome sample processing, the three solutions of A, B, and C in the kit and the mass spectrometer water were prepared according to the formula of 1:1:1:6 to make a mixed solution, and finally 1 part of D solution was added, mixed evenly, and prepared Complete treatment solution.

[0060] Samples of this example: gastric cancer tissue and adjacent normal tissue

[0061] The operation is as follows:

[0062]1. Weigh the empty EP tube and mark it. Each pair of cancer tissue (Tumor, T) and paired adjacent normal tissue (Nearby tissue, N) samples were guaranteed to be processed in parallel at the same time. After the tissue samples were taken out from -80°C, they were f...

Embodiment 3

[0070] The kit of this embodiment contains four solutions A, B, C, and D, and the ingredients are as follows in Table 3:

[0071] table 3

[0072] Solution name Element A Tris-HCl pH=8.8 B 400mM CAA solution C 100mM TCEP solution D 10% (W / V) DOC solution

[0073] Before proteome sample processing, the three solutions of A, B, and C in the kit and the mass spectrometer water were prepared according to the formula of 1:1:1:6 to make a mixed solution, and finally 1 part of D solution was added, mixed evenly, and prepared Complete treatment solution.

[0074] Samples of this example: gastric cancer tissue and adjacent normal tissue

[0075] The operation is as follows:

[0076] 1. Weigh the empty EP tube and mark it. Each pair of cancer tissue (Tumor, T) and paired adjacent normal tissue (Nearby tissue, N) samples were guaranteed to be processed in parallel at the same time. After the tissue samples were taken out from -80°C, they were ...

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Abstract

The invention discloses a proteome sample processing kit box and processing method and an application of the processing method. The kit box comprises a tri(2-carbonyl ethyl) phosphorus hydrochloride,2,2-dichloro acetamide, sodium deoxycholate and a buffer solution. The application of the processing method provided by the invention is as follows: before a protein sample is processed, the tri(2-carbonyl ethyl) phosphorus hydrochloride, 2,2-dichloro acetamide, sodium deoxycholate and buffer solution in the kit box are prepared into treating fluid, wherein splitting and whole protein degenerationin cells are performed on cell / tissue samples via the sodium deoxycholate, the tri(2-carbonyl ethyl) phosphorus hydrochloride is used as a reducing agent, the 2,2-dichloro acetamide is used as an alkylate reagent, and splitting and reductive alkylation of the proteome sample can be completed by several minutes in the treating fluid. That is to say, according to the kit box, processing method andapplication, the splitting and reductive alkylation are completed in one step, so that the proteome sample treating process is simple, and the time is saved.

Description

technical field [0001] The present invention relates to the technical field of proteome detection, in particular, to a proteome sample processing kit, processing method and application. Background technique [0002] In recent years, mass spectrometry technology has developed rapidly. Mass spectrometry companies offer mass spectrometers with ultra-high resolution and with multiple detectors. The use of this mass spectrometer in conjunction with high-pressure liquid phase greatly improves the detection efficiency of proteomics based on mass spectrometry, making it possible to construct large-scale clinical sample proteomic data sets. [0003] At present, the preparation of mass spectrometry and liquid chromatography samples in the laboratory widely uses 8M urea to cleavage proteins, supplemented with 10mM M DTT (dithiothreitol), 10mM IAA (iodoacetamide) reduction, alkylation and digestion to obtain peptide samples. In a typical operation, the first sRP liquid phase separatio...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88G01N2030/8831
Inventor 秦钧汪宜丁琛宋雷刘明伟杨星
Owner 北京谷海天目生物医学科技有限公司
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