This service is used to detect post-translational modifications (PTMs) on proteins. The service can be applied to purified proteins, recombinant proteins or proteins in complex mixtures. Samples should be submitted as gel bands. The range of modifications is limited to those readily identifiable in a database search, such as methylation of lysine or arginine, ubiquitination of lysine, deamidation of glutamine or asparagine, glycation of lysine, phosphorylation of serine, threonine or tyrosine (without enrichment), or acetylation of lysine.
In more complex samples, fractionation can be employed, the extent of the fractionation correlates with the number of proteins and their PTMs that are detected. The greater the number of fractions, the greater the number of PTMs identified.
Following trypsin digestion the peptide mixture is analyzed by LC/MS/MS. The peptides are fragmented in the mass spectrometer to yield diagnostic patterns that can be matched to protein sequence databases via computer algorithms.
A 10 fraction analysis is suitable for the analysis of low complexity samples e.g. HPLC fractions, semi-purified extracts. A 20 fraction analysis is for medium complexity samples e.g. subcellular fractions, pooled HPLC fractions. A 40 fraction analysis is suitable for complex samples e.g. whole cell lysates, whole tissue extracts.
Sample types: 2D gel spots, SDS-PAGE bands, recombinant proteins, purified proteins, cell lysates, tissue homogenates, biofluids, FFPE extracts
Data turn around: Two weeks or less
Report format: PDF report