LC-MS Detection of Phosphorylation Sites
In the past, the most common method for determining phosphorylation sites was stepwise chemical degradation, such as serine and threonine phosphorylation analysis methods and tyrosine phosphorylation analysis methods. With the advancement of life science technology, chemical methods have been replaced by mass spectrometry methods. Currently, phosphorylation sites can be detected by liquid chromatography-mass spectrometry (LC-MS).
Liquid chromatography-mass spectrometry generally consists of a front-end liquid chromatography and a mass spectrometer. The liquid chromatography serves as the separation system, and the mass spectrometer serves as the detection system. The sample is separated from the mobile phase in the mass spectrometry part, ionized, and the ion fragments are separated by mass number by the mass analyzer of the mass spectrometer, and a mass spectrum is obtained through the detector. Large-scale analysis of phosphorylated proteins in complex protein mixtures includes the identification of phosphorylated proteins/phosphorylated peptides, as well as sequencing of phosphorylated residues and identification of phosphorylation sites, all of which can be detected by LC-MS methods.
LC-MS detection analysis is a useful tool for identifying specific phosphorylated proteins/peptides and locating specific phosphorylated residues. It has high sensitivity and high resolution and can identify single proteins or peptides. Although the signal of phosphorylated peptides is usually weaker than that of non-phosphorylated peptides, the latest technology can now enrich mass spectrometry signals, including immobilized metal affinity chromatography, phosphorylation-specific antibody enrichment, chemical modification methods, and partial replacement of phosphate groups with biotin.
Biotech company Biotye adopts Thermo Fisher's Q Exactive HF mass spectrometry platform, Orbitrap Fusion mass spectrometry platform, and Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC to launchphosphorylation quantitative proteomics analysisservice package. You only need to inform us of your experimental objectives and send the samples. Biotye is responsible for the subsequent project tasks, including protein extraction, protein digestion, enrichment of phosphorylated peptides, peptide separation, mass spectrometry analysis, raw data analysis, and bioinformatics analysis.
Related services:
Multipathway phosphoproteomics
Post-translational modification proteomics analysis
Histone post-translational modification analysis
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