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SILAC QUANTITATIVE PROTEOMICS (SILAQTM)

SILAQTM service comprises an advanced workflow that can be customised to the specific needs of the client and their project, including project design, implementation through wet lab experiments, high-resolution mass spectrometry, data processing, analysis and interpretation and delivery of a final project and data report.

SILAQTM is used in high throughput, quantitative analyses of cellular proteomes, large protein complexes and protein-protein interactions. The technology involves the use of Stable Isotope Labelling with Amino acids of Cells in culture (SILAC) technique combined with high sensitivity mass spectrometry and specialised software for data processing and analysis. Our SILAQTM approach provides a quantitative and unbiased strategy that not only identifies proteins, but also simultaneously annotates their properties and interactions.

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POST-TRANSLATIONAL MODIFICATIONS PROTEOMICS

1. Phosphoproteomics and phosphosite mapping

Protein phosphorylation is the most important reversible post-translational modification that occurs in cells. Protein kinases and phosphatases add or remove, respectively, phosphate groups that regulate enzymatic activity, protein-protein interactions and protein tertiary structures. Knowledge on how phosphorylation affects the behaviour of proteins in the cell is essential in understanding signal transduction pathways. Identification of phosphorylation sites in a protein could be very challenging since most phospho-proteins contain sub-stoichiometric sites of phosphorylation.

DCP uses multidimensional MS-based strategies that allow for selective and sensitive detection and identification of multisite phosphorylation on proteins. The identification strategy includes:

  • Global changes in phosphoproteome
  • Phosphopeptide detection
  • Phosphopeptide identification
  • Phosphopeptide sequencing

2. Methylation proteomics

3. Acetylation proteomics

4. Ubiquitination proteomics

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OTHER QUANTITATIVE PROTEOMICS

Metabolic labeling is not always appropriate or is difficult to use for some quantitative proteomics analyses e.g. using human or animal tissues, patient samples, serum and primary differentiated cells that are not able to undergo multiple cell division cycles in culture. Other quantitative proteomics services available at Dundee Cell Proteomics include:

  • Isobaric labeling using: tandem mass tags (TMT) or isobaric tags for relative and absolute quantitation (iTRAQ)
  • Label-free quantitation
  • Isotope-coded affinity tag (ICAT)

We will discuss projects with clients and then determine the appropriate service that will provide a solution to their research and development problem.

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