Stable Isotope Labeling of Cell Culture with Amino Acids (SILAC) .
Stable Isotope Labeling with Amino Acids (SILAC) of cell culture is a highly qualified method to identify and quantify changes in protein samples. Within quantitative proteomics, cell samples are differentially labeled with non-radioactive isotope labels.
Studies are performed using cells with normal isotopes 12C and 14N. When mammalian cells are grown using SILAC reagents, the proteins become substituted with the heavy forms of arginine (R) and lysine (K). The amino acids arginine and lysine are replaced with isotopes 13C and/or 15N. The replacement concerns the sites of trypsin cleavage. This generates a set of tryptic peptides. Mass spectrometry accelerates, detects and quantifies the changes in mass in proteins under investigation. Identification and quantification of relative differences in proteins abundance as low as 30% between treated and normal cells are possible.
Create conveniently sets of tryptic peptides, each with increased mass that can readily be detected and quantificated by mass spectrometry. As SILAC media only contain heavy amino acids, only heavy amino acids can be incorporated. The share of SILAC proteins are compared after cell stimuli relative to standard proteins prior to stimuli. In case certain proteins are "heavier" one can conclude that they have been augmented by the stimulus. In the reverse case, stimuli seem to have reduced certain proteins.
PRIMACYT cooperates with Dundee Cell Products Ltd in Dundee/UK and is the Authorized Distributor for • Germany, Austria, Switzerland, • France, Belgium, Netherlands, Luxembourg. • Poland and the Czech Republic.
Move forward to quantitative high throughput analysis of large protein complexes and interactions between proteins.
Quantitative analysis of large protein complexes and interactions between proteins.
Protein expression profiling.
Discover how inhibitors or other perturbations affect the dynamic properties and/or cellular distributions of proteins.
Identify specific interaction partners of proteins under study.
Functional characterization of protein components in cell-based assays.
Drug discovery and development
Systematic evaluation of the effects of compounds on proteins. Improved characterization of chemical compound mechanisms and specificity. Identification of potential side effects of novel hit and lead compounds prior to clinical trials.
Utilize the benefits and advantages of quantitative protein analysis and identification.
Analysis of post-translational modifications and low abundance proteins. Analysis of relative changes in proteins abundance from different cell treatments. Analysis of phosphoproteins, membrane proteins, and gene knockdown experiments. Analysis of proteins without availability of antibodies. Expression analysis and profiling of normal cells compared to diseased cells.