Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for relative quantitative analysis of protein phosphorylation modification level

A quantitative analysis and phosphorylation technology, applied in the field of analysis, can solve the problems of large labeling reagents, increased experimental cost and complexity, consumption, etc., to achieve the effect of improving sensitivity and accuracy, reducing experimental cost, and reducing operational complexity.

Inactive Publication Date: 2013-06-19
TIANJIN INT JOINT ACADEMY OF BIOTECH & MEDICINE +1
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this isotope-labeled quantitative mass spectrometry method needs to consume a large amount of labeled reagents, resulting in a significant increase in experimental costs and operational complexity.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for relative quantitative analysis of protein phosphorylation modification level
  • Method for relative quantitative analysis of protein phosphorylation modification level

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0028] The following examples can enable those skilled in the art to understand the present invention more fully, but do not limit the present invention in any way.

[0029] Experimental samples: mutant (mut) and wild type (wt) of a certain protein kinase

[0030] experimental method:

[0031] 1. The two experimental samples mut and wt were digested with trypsin to convert the protein into a polypeptide mixture.

[0032] 2. Add an appropriate amount of internal standard substance (enzymatic hydrolysis product of β-casein) to the enzymatic hydrolysis samples of the two proteins to enrich the phosphorylated peptides together.

[0033] 3. Equilibrate the titanium dioxide affinity column with solution A (80% acetonitrile, 0.4% trifluoroacetic acid) and solution B (25% lactic acid, 60% acetonitrile, 0.3% trifluoroacetic acid) respectively, then mix the enzymolysis sample with Solution B is loaded onto the affinity column together. During centrifugation, the phosphorylated peptide...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides an isotope label-free method for relative quantitative analysis of a protein phosphorylation modification level. The method comprises: conducting enzymolysis on protein to make it into peptide fragments, making use of a data-independent MS / MS technique to determine peptide fragments with phosphorylation sites, and utilizing a label-free quantitation technique to carry out relative quantitative analysis on the peptide fragments with phosphorylation sites. Compared with the existing methods combining data-dependent MS / MS analysis and stable isotope-labeling technologies, the method does not need an isotope labeling reagent, thus having the advantages of reducing the experiment cost and operation complexity.

Description

technical field [0001] The invention relates to an analysis method, in particular, the invention relates to a method for relative quantitative analysis of protein phosphorylation modification level. Background technique [0002] With the completion of the International Human Genome Project in June 2000, human beings have entered the post-genome era from the genome era. In the post-genome era, the main research object of life sciences is functional genomics, and the function of genes depends on the executors of their functions. -Protein is implemented, so the research on protein expression, location, modification state and protein interaction has become a hot research field in the post-genome era. [0003] In the living body, proteins need to undergo gene transcription, translation, and post-translational modification before they can function, and are transported to specific parts of cells or tissues to function. Most proteins are inactive before post-translational modificat...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N1/40G01N27/62G01N30/08
Inventor 水雯菁杨诚刘丹魏晓超徐金华
Owner TIANJIN INT JOINT ACADEMY OF BIOTECH & MEDICINE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com