What can SILAC tell you?
SILAC has emerged as a very powerful method to study cell signaling, post translation modifications such as phosphorylation, protein–protein interaction and regulation of gene expression. It can be used to distinguish between proteins secreted by cells in culture and serum contaminants.
Is SILAC quantitative?
SILAC. Stable Isotope Labeling with Amino acids in Cell culture (SILAC) is a quantitative proteomic approach which allows the comparison of protein levels in up to 3 cultured cell extracts. With this approach it is possible to identify and quantitate thousands of proteins in a single experiment.
How does SILAC MS work?
In SILAC experiments, proteins are metabolically labeled by culturing cells in media containing normal and heavy isotope amino acids. This makes proteins from the light and heavy cells distinguishable by mass spectrometry (MS) after the cell lysates are mixed and the proteins separated and/or enriched.
How does SILAC differ from iTRAQ?
The main differences among labeling techniques are (i) SILAC and ICAT labeling are applied on intact proteins, while iTRAQ labeling is performed on peptides, and (ii) in the case of SILAC and ICAT, peptides are quantified during MS analysis, while in the case of iTRAQ, quantitation occurs during fragmentation, i.e., MS …
What is SILAC technique?
SILAC (stable isotope labeling by amino acids in cell culture) is a technique based on mass spectrometry that detects differences in protein abundance among samples using non-radioactive isotopic labeling.
What does SILAC mean?
Stable isotope labeling by amino acids in cell culture (SILAC) is a multiplexing quantitative proteomic method that uses labeled isotopically heavy amino acids, for example 13C6,15N2-lysine and 13C6,15N4-arginine, incorporated metabolically into the whole proteome [102,103].
Who invented SILAC?
Made possible also by recent advances in instrumentation and bioinformatics analysis, several noteworthy proteomics studies have been reported over just the last year, in particular using metabolic labeling techniques such as the SILAC (stable isotope labeling by amino acids in cell culture) method invented by Matthias …
What is the difference between iTRAQ and TMT?
iTRAQ and TMT are chemical labels which are reacted with complex protein samples after proteolysis to peptides. iTRAQ (isobaric tagging for relative and absolute quantification) is available in 4-plex and 8-plex formats, while TMT (tandem mass tags) is available in 2-plex, 6-plex and (since recently) 10-plex formats.
How does Tandem Mass Tag work?
Isobaric tags for relative and absolute quantitation (iTRAQ) or Tandem Mass Tags (TMT) is a quantitation method in which peptide N-terminus and side chain amines are covalently labeled with tags of varying masses. Up to 16 different samples or treatments can be analyzed using this technique.
What is TMT technique?
From Wikipedia, the free encyclopedia. A tandem mass tag (TMT) is a chemical label that facilitates sample multiplexing in mass spectrometry (MS)-based quantification and identification of biological macromolecules such as proteins, peptides and nucleic acids.
How does TMT Labelling work?
How TMT quantitative proteomics works. Because the isobaric tags possess the same chemical properties, all peptides from different TMT-labeled samples co-elute during LC separation. Once the peptides enter the mass spectrometer, they are detected simultaneously as a single and indistinguishable precursor ion peak.