SILAC kits and reagents for mouse embryonic stem cell proteomic analysis.

The Thermo Scientific Pierce Mouse Embryonic Stem Cell SILAC Kit, Media and Reagents are specifically designed for analysis of protein expression by mass spectrometry using a method called stable isotope labeling with amino acids in cell culture (SILAC). The kit includes a complete set of specialized media, supplements and heavy and light isotopes of lysine for performing a complete experiment. In addition, individual components are offered, including the expansion DMEM and low-osmolarity DMEM (media), compatible serum and serum-substitute supplements and alternative lysine and arginine isotope pairs.

What is 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. The SILAC Method uses in vivo metabolic incorporation of "heavy" ¹³C- or ¹⁵N-labeled amino acids into proteins followed by mass spectrometry (MS) analysis for accelerated comprehensive identification, characterization and quantitation of proteins.

... Learn more about the SILAC method.

Highlights of SILAC Products for Mouse Embryonic Stem Cells:

• Optimized, specialized media – the Expansion DMEM and Low-Osmolarity DMEM are specifically designed and qualified for mouse embryonic stem cell propagation and SILAC protein expression profiling with mass spectrometry
• High-quality supplements – the Stem Cell Screened Dialyzed FBS and Serum Substitute for Mouse Embryonic Stem Cells are tested to ensure that they do not promote stem cell differentiation; the Phenol Red-Free MEM Media for SILAC can be used with estrogen responsive cells such as the breast adenocarcinoma cell line MCF7, without inducing an artificial estrogen response
• Flexible and compatible – media and the SILAC procedure are compatible with all standard mass spectrometry instrumentation
• Multiplex capabilities – several alternative isotopes of arginine and lysine are available that allow for analysis of multiple growth conditions in each experiment. (For additional individual SILAC reagents and amino acid isotopes see related SILAC Product page).
• Thousands of proteins – in vivo labeling with arginine and lysine isotopes using endogenous translational machinery enables expression analysis of the entire proteome; investigate any protein or group of proteins you choose

SILAC Applications:

• 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

Proteome analysis of GRP94 -/- mouse embryonic stem cells using SILAC and muscle differentiation conditions. A. Protein expression changes observed with SILAC between GRP94 -/- mouse embryonic stem cells under growth (13C6, 15N2-"heavy" lysine) versus muscle differentiation conditions ("light" lysine, 0.8% DMSO for six days). B. Proteins identified and quantified after analysis with a Thermo Scientific LTQ-FT Mass Spectrometers and Bioworks 3.3.1 Software, and MAPPFinder. C. Significantly regulated gene ontologies were observed in GRP94 -/- mouse embryonic stem cells under muscle differentiation conditions. These cells cannot differentiate into muscle without GRP94 function or insulin-like growth factor supplementation. The down regulation of nearly 30 ribosomal proteins during "frustrated" muscle differentiation indicates that protein translational machinery is being profoundly regulated.

Read the following Poster Presentation:

• SILAC studies of murine embryonic stem cell differentiation into muscle

SILAC Literature References

1. Amanchy, R. et al. (2005). Science STKE 267: 1-20
2. Blagoev, B., et al. (2004). Nat Biotechnol 22(9): 1139-1145.
3. Everley, P. et al. as presented at The American Society for Biocehmistry and Molecular Biology. MCP Papers in Press. Published on July 11, 2007 as Manuscript M700057-MCP200.
4. Kratchmarova, I., et al. (2005). Science 308(5727): 1472-1477.
5. Mann, M. (2006). Nat Rev Mol Cell Biol 7(12): 952-958.
6. Ong, S. E., et al. (2002). Mol Cell Proteomics 1(5): 376-386.
7. Selbach, M. and Mann, M. (2006). Nat. Methods 3(12): 981-3

Related Products and Links
Individual SILAC Reagents – including universal supplements and additional amino acid isotopes
Standard SILAC Kits – including DMEM and RPMI-160 media for SILAC
AQUA and Heavy Peptides – custom peptide isotopes for mass spectrometry
Silver Stain for Mass Spectrometry – exceptional sensitivity and recoverability for MS
In-Gel Trypsin Digest Kit – optimized kit for protein MS sample preparation
Prepare SILAC peptides using the In-Gel Tryptic Digestion Kit (Tech Tip #60)

The purchase of this product conveys a non-transferable license to the Purchaser to use this product in methods protected under U.S Patent 6,653,076 (owned by University of Washington) for research purposes only.