![]() Procedure summary for MS experiments with TMT Isobaric Mass Tagging Reagents PreOmics iST-NHS Kit, a fast, reliable, reproducible sample preparation for chemical labeling (iTRAQ and TMT) There are multiple different kits available (2-plex, 6-plex, 10-plex, 16-plex and 18-plex), including bulk, so you can adjust your order to your specific experimental needs. Of that peptide from each biological sample determined from the areas of each respective reporter ion peak. Upon fragmentation, the reporter ion can be clearly distinguished, and the identity of the peptide determined from the sequence of the larger MS/MS peptide fragments, and the quantity Regardless of which reporter ion is attached. Since all versions of the reagent tag are identical in molecular weight, the same peptides originating from different samples will have the same mass in MS space ![]() “reporter ion” region and “balance mass” region within the compound. All versions of each tag have the same molecular mass but the positions of heavy and light isotopes are adjusted in order to affect the mass of a Tandem Mass Tags (TMT and iTRAQ) are isobaric multiplexing tags. Labeled peptides, which increases the complexity in the MS space and limits to comparing only 2 or 3 sample types. Isotope coded affinity tags ICAT and Stable Isotope Labeling by Amino Acids in Cell Culture, or SILAC, are technologies that provide a mass difference in the molecular weights of differentially Much of the bias that can be introduced when comparing peaks between different experiments since the data from all samples are collected within the same experiment. Quantitation is performed on the pairs of peaks in the MS data and identification is performed using the MS/MS fragment data. ![]() Identical compounds from the different samples co-elute as pairs of peaks and can be distinguished by the mass differenceīetween the heavy and light isotope labels. The samples are then mixed together and analyzed in the same experiment. With stable isotope labeling, one sample is derivatized with a “light” version of a chemical tag while another sample is labeled with a version of the same tag that incorporates a “heavy” isotope. Stable Isotope Labeling Strategies Protein Quantitation using Mass Spectrometry ![]()
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