SILAC-based Proteomics Analysis

SILAC-based quantitative proteomics (SILAQ) is an innovative technology used in high

throughput quantitative analysis of large protein complexes, protein-protein and protein-small
molecule interactions. SILAC service of Creative Proteomics provides an unbiased strategy
that can reveal how specifically either inhibitors, or other perturbations, affect the dynamic
properties and cellular distributions of proteins. It can also be used as a sensitive and
effective method to determine the specific interaction partners of proteins in the cell.

SILAC Workflow:

SILAC relies on metabolic incorporation of a given 'light' or 'heavy' form of the amino acid into
two samples. As the two isotopically labeled amino acids are essentially chemically identical,
their incorporation does not interfere with normal cell growth, while leading to
proteins/peptides that are distinguishable by mass and thus are ideal for mass spectrometric
analysis. The method relies on the incorporation of amino acids with substituted stable
isotopic nuclei. Thus in an experiment, two cell populations are grown in culture media that
are identical except that one of them contains a 'light' and the other a 'heavy' form of a
particular amino acid (e. g. 12C and 13C labeled L-lysine, respectively). The SILAC samples
are then subjected to enzymatic digestion and LC/MS analysis. The protein quantification is
carried out on the protein level. In addition, SILAC approaches are well suited for monitoring
 changes in post-translational modifications. Examples for these applications include the
measurement of changes in protein phosphorylation and methylation.
Advantage of our SILAC technique:
 Cutting-edge facilities & optimized protocols
 Harmless in vivo labeling
 Compare up to three samples at one time
 High sensitivity
 Suitable for study of protein-protein-interaction
 Post-translational modification is detectable
Bioinformatic Analysis:
 Functional annotation and enrichment analysis
 Clustering analysis
 Network analysis
 Statistical analysis