SILAC Proteomics: Innovative Tech for Quantitative Proteomics & Interaction Analysis, Essays (university) of Biology

Silac proteomics is a cutting-edge technology used for high-throughput analysis of large protein complexes, interactions, and modifications. It relies on metabolic incorporation of 'light' or 'heavy' isotopically labeled amino acids, enabling mass spectrometric analysis and protein quantification. Silac offers advantages such as harmless in vivo labeling, comparison of up to three samples, high sensitivity, and detection of post-translational modifications. Creative proteomics provides silac services with optimized protocols and bioinformatic analysis tools for functional annotation, clustering, network analysis, and statistical analysis.

Typology: Essays (university)

2018/2019

Uploaded on 10/15/2019

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SILAC 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.
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SILAC 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 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.