Birdshot Proteomics Sequencing

Protein sequencing is the determination of the primary structure of proteins, which is the sequence of amino acids that make up the protein. The primary structure of a protein largely determines its spatial structure and biological functions. Currently, commonly used protein sequence analysis methods mainly include terminal sequencing methods (such as Edman degradation method and carboxypeptidase method) and mass spectrometry-based methods. Shotgun is a mass spectrometry-based protein sequencing method. Based on the "bottom-up" mass spectrometry analysis strategy, through the data-dependent acquisition (DDA) mode, it aims to achieve unbiased identification and quantification of proteins in biological samples.

 

The general process of shotgun protein sequencing is to first digest the protein sample into peptides, and then separate the peptides by high-performance liquid chromatography (HPLC). The separated peptides directly enter the mass spectrometer for analysis. Mass spectrometry analysis includes two processes, primary mass spectrometry (MS1) and secondary mass spectrometry (MS2). The primary mass spectrometry obtains the mass-to-charge ratio (m/z) and intensity of the peptides eluted in the HPLC MS1; the secondary mass spectrometry determines the peptide sequence, MS2 selects a single peptide to fragment it to obtain the spectral data of the peptide. The retrieval software compares the secondary mass spectrometry information with the corresponding database, combines the scoring of matching degree and mismatch filtering, to obtain the exact sequence of the peptide, and then concatenate the complete sequence of each protein, thereby realizing the identification of the protein.

 

Service Advantages

1. High Throughput

Shotgun protein sequencing can simultaneously identify and quantify thousands of proteins, greatly improving the efficiency of proteomics research.

 

2. High Sensitivity

This technology can detect low-abundance proteins, providing a powerful tool for discovering new biomarkers and studying small changes in proteins.

 

3. Wide Range of Applications

Shotgun protein sequencing is suitable for different types of samples, including body fluids, tissues, and cells, providing convenience for various biological research.

 

MtoZ Biolabs uses Thermo Fisher's Orbitrap Fusion Lumos mass spectrometry platform combined with nanoLC-MS/MS nanoscale chromatography to provide you with an efficient and accurate shotgun proteomics sequencing analysis service one-stop technical package. It can perform terminal sequencing, full-length sequencing, and de novo sequencing of various proteins, and can accurately determine proteins with N-terminal modifications and newly discovered protein sequences. Free project evaluation!