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 Features of SILAC Protein Quantitation Kit – RPMI 1640: 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