Protein digestion in-gel or in-solution using a proteolytic enzyme.
When proteins interact with other proteins, they form protein-protein interactions (PPI). PPIs are fundamental for the function of cells and consequently any living organism. Gene expression, cell growth, proliferation, apoptosis, and intercellular communication are among the many biological processes that are facilitated by proteins. Samples are lysed, affinity-purified and digested into peptides using trypsin, followed by C18-desalting. Purified samples are subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis.
Proteomes are sets of proteins produced in an organism, system, or biological context. For example, one can refer to the proteome of a species, e.g. the human proteome, or the proteome of an organ, e.g. the liver. Besides overall understanding of human biology, proteomics has created opportunities to study proteins in diseases. Disease proteomics aim to understand how altered protein expression, structure and function can cause illness.
Depending on the specific needs of the project, the starting point for phosphoproteomics analysis can be a piece of tissue, cell pellet or a lysate of known protein concentration. The sample is phosphoenriched with our in-house enrichment protocol, proteolytically digested to peptides, desalted and analyzed on a LC-MS/MS.