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

Description

• Features of the SILAC Protein Quantitation Kit, RPMI 1640, include: • Efficient—up to 100% label incorporation into proteins of living cells • Reproducible—minimizes 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 help ensure that it is sterile, endotoxin-free, and cell culture compatible Applications of SILAC Protein Quantitation Kit: • 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 to study normal vs disease cells • Identification and quantification of hundreds to thousands of proteins in a single experiment 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' 13 C- or 15 N-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
• Trypsin SILAC kits are supplied with 13 C 6 15 N<sub2 L-lysine-2HCl and 13 C 6 15 N 4 L-arginine-HCl to label and quantify lysine-containing and arginine-containing peptides from trypsin protein digests
• Each kit also includes 2 x 500 mL of medium and dialyzed serum
• When combined with Thermo Scientific protein/peptide sample enrichment products, 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.