Thermo Scientific™ SILAC Protein Quantitation Kit (LysC), RPMI 1640

Description

Thermo Scientific SILAC Protein Quantitation Kit – RPMI 1640 is used for the specific analysis of protein expression by mass spectrometry using stable isotope labeling with amino acids in cell culture (SILAC).

Stable isotope labeling with amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. This approach entails the in vivo metabolic incorporation of “heavy” 13C- or 15N-labeled amino acids into proteins followed by mass spectrometry (MS) for the accelerated and comprehensive identification, characterization and quantitation of proteins.

Because peptides labeled with heavy and light amino acids are chemically identical, they co-elute during reverse-phase column pre-fractionation and therefore are detected simultaneously during MS analysis. The relative peak intensities of multiple isotopically distinct peptides from each protein are then used to determine the average change in protein abundance in the treated sample.

Each kit includes all necessary reagents to isotopically label cells with 13C6 L-lysine-2HCl, including media, heavy and light amino acids and dialyzed serum. Heavy L-arginine-HCl derivatives (Part No. 88210, 89990) are available separately and can be combined with Pierce SILAC Protein Quantitation Kits to enhance peptide isotope label coverage. Kits are compatible with mammalian cell lines adapted to grow in either DMEM or RPMI 1640 media. Kits with specialized media formulations are also available for human and murine stem cell lines. When combined with Thermo Scientific Protein/Peptide Sample Enrichment Products, Pierce SILAC Protein Quantitation Kits also enable MS analysis of low-abundance proteins such as cell surface proteins, organelle-specific proteins and post-translational protein modifications such as phosphorylation or glycosylation.

Recommended for: Quantitative analysis of relative changes in protein abundance from different cell treatments; Quantitative analysis of proteins for which there are no antibodies available; Protein expression profiling of normal vs. disease cells; Identification and quantification of hundreds to thousands of proteins in a single experiment

• Efficient—100% label incorporation into proteins of living cells
• Reproducible—eliminates intra-experimental variability caused by differential sample preparation
• Flexible—media deficient in both L-lysine and L-arginine, allowing for more complete proteome coverage through dual amino acid isotope labeling
• Compatible—label proteins expressed in a wide variety of mammalian cell lines adapted to grow in RPMI 1640 medium, including HeLa, 293T, COS7, U2OS, A549, A431, HepG2, NIH 3T3, Jurkat and others
• High-quality supplements—heavy amino acids with >99% isotope purity; dialyzed FBS tested to ensure that it is sterile, endotoxin-free, and cell culture compatible