Quantification of proteins using isobaric labeling (tandem mass tag or TMT) starts with the reduction of disulfide bonds in proteins with Dithiothreitol (DTT). Alkylation with iodoacetamide (IAA) after cystine reduction results in the covalent addition of a carbamidomethyl group that prevents the formation of disulfide bonds. Then, overnight digestion of the proteins using trypsin or trypsin/LyC mixture, Tandem Mass Tag (TMT) labeling on Lysine residues, pooling of all samples and the fractionation of peptide mixture.
Related Posts
-
Spatial gene expression data analysis on Cluster : 10X Genomics, Space Ranger
Running spaceranger as cluster mode that uses Sun Grid Engine (SGE) as queuing. There are -
Taxonomic and functional profiling of the microbiome – whole genome shotgun metagenomics
This workflow consists of taxonomic and functional profiling of shotgun metagenomics sequencing (MGS) reads using -
Taxonomic and diversity profiling of the microbiome : 16S rRNA gene amplicon sequence data
The 16S ribosomal RNA (rRNA) gene of Bacteria codes for the RNA component of the