At present, the identification of protein gel spots and strips is the most widely used means of proteomics research. Protein mixtures are separated by two-dimensional gel electrophoresis (2DE) and SDS-PAGE, and target protein spots and strips are collected. After in-gel enzymolysis, mass spectrometry is used to determine the protein composition of the sample. The basic principle is to identify the peptides and protein types using the ion peak distribution of secondary mass spectrometry obtained by the mass spectrometer.
Figure 1. 2DE gel images. ( Zhang N.,et al., 2016)
CD Mitochondria combines MALDI-TOF mass spectrometry system and LC-ESI-MS/MS mass spectrometry system to provide professional and accurate protein identification services for protein glue spots, strips, and IP samples, and a series of optimization of in-gel enzymolysis, which significantly improves the success rate of mass spectrometry identification and peptide coverage of protein in-gel enzymolysis.
The identifiable samples include protein strips isolated from 1D-SDS-PAGE, protein spots in 2DE-PAGE, and protein expression products. Among them, the pre-treatment process of protein sample identification is carried out strictly following the method published by Nature Methods to ensure a higher peptide coverage in the identification test. The principle of mass spectrometry identification is based on shot-gun. Protein bands or protein solutions are digested into peptide mixtures, and some relatively simple components are separated by high-performance liquid chromatography and analyzed by the high-resolution mass spectrometer. According to the secondary mass spectrometry information and the corresponding database comparison, combined with matching degree scoring and mismatch filtering, the retrieval software can get the exact sequence of the peptide, and then splice the complete sequence of each protein to identify the protein.
CD Mitochondria will provide you with the experimental steps, relevant MS parameters, MS images, raw data, and protein details immediately after the end of the experiment. CD Mitochondria always provides you with customized services for mitochondrial research, and our one-stop research and analysis platform can meet all your research needs.
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