lc-ms/ms protein identification
LC-MS/MS (Liquid Chromatography-Tandem Mass Spectrometry) protein identification is an advanced analytical technique widely used in proteomics research. This method combines high-performance liquid chromatography (LC) separation technology with highly sensitive tandem mass spectrometry (MS/MS) detection technology, enabling the identification and quantitative analysis of proteins in complex samples. Below is the basic process and principle of LC-MS/MS protein identification.
I. Basic Process
1. Sample Preparation:
First, proteins need to be extracted from the sample and hydrolyzed into shorter peptides through enzymatic digestion (commonly using trypsin).
2. Liquid Chromatography Separation:
The processed sample is separated in the chromatographic column using a liquid chromatography system based on the hydrophilicity or other chemical properties of the peptides. This step effectively reduces sample complexity and enhances the resolution and sensitivity of subsequent mass spectrometry analysis.
3. Mass Spectrometry Detection:
The separated peptides are introduced into the mass spectrometer for detection. The mass spectrometer first measures the mass-to-charge ratio (m/z) of the peptides, then selects specific peptides for secondary mass spectrometry (MS/MS) analysis, where the peptides are further fragmented and the m/z values of the fragments are measured.
4. Data Analysis:
Software is used to analyze the MS/MS data. By comparing the mass-to-charge ratio information of the peptides and their fragments with protein sequences in a database, the peptides can be identified. The identified peptides allow for the inference of the identity of proteins in the original sample.
II. Applications and Advantages
1. Protein Identification:
Capable of identifying proteins in unknown samples, including low-abundance proteins.
2. Protein Quantification:
Specific quantification strategies (such as labeling or label-free methods) can be used to quantitatively analyze proteins in a sample.
3. Post-translational Modification Analysis:
Can be used to study post-translational modifications of proteins, such as phosphorylation and glycosylation.
LC-MS/MS protein identification is a highly sensitive and accurate method that provides extensive information about protein molecules, which is essential for a deep understanding of the composition and function of proteins in biological systems.
BiotechPack, A Biopharmaceutical Characterization and Multi-Omics Mass Spectrometry (MS) Services Provider
Related Services:
Protein Tandem Mass Spectrometry Identification
Protein Mass Spectrometry Identification
Protein Identification from Gel Spots, Gel Strips, IP Samples
Analysis Services Based on Liquid Chromatography (LC)
Post-translational Modification Proteomics Analysis
Protein Purity Analysis (Size Exclusion/Reverse Phase Chromatography)
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