Stable isotope labeling in cell culture

Stable Isotope Labeling in Cell Culture (SILAC)... 

Gronborg, M., IwahorrA., Pandey, A. (2003). A proteomic approach for quantification of phosphorylation using stable isotope labeling in cell culture. Anal. Chem. 75, 6043-6049. 

Egorova-Zachemyuk TA, Bosman GJ, DeGrip WJ (2011) Uniform stable-isotope labeling in mammalian cells formulation of a cost-effective culture medium. Appl Microbiol Biotechnol 89 397 6... 

The approach recruited to chemical proteomics in Reference [17] is called SILAC (stable isotope labeling with amino acids in cell culture) and is important in comparative proteomics (Figure 1). SILAC works well with cultured mammalian cells, but prokaryotes defeat it by metabolizing the label (usually supplied in lysine and arginine) into other amino acids. For applications beyond cultured eukaryotic cells, the reductive methylation route to differential labeling [18] is among the alternatives [15]-... 

Guerrero et al. (2006) used this technique along with the quantitative mass spec strategy called SILAC (stable isotope labeling of amino acids in cell culture Ong et al., 2002) to identify the yeast proteins that interact with the 26 S proteasome. 

Ong, S.-E., Blagoev, B., Kratchmarova, I., Kristensen, D.B., Steen, H., Pandey, A., and Mann, M. (2002) Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics. Mol. Cell. Proteomics 1, 376-386. 

Figure 4.10. Proteomic analysis by SILAC. Proteomic analysis by SILAC or stable isotope labeling of amino acids in cell culture utilize de novo metabolic incorporation of stable-isotope-labeled amino acids during protein synthesis. Cells can be cultured with various combinations of stable-isotope-labeled amino acids such as lysine or arginine. Tyrosine has been used in phosphoprotein studies of tyrosine residues. About five or six cell divisions are needed for complete labeling of proteins in cell cultures prior to experimentation. Labeled cells from control and treatment(s) lysates are combined and digested. Quantitation and identification are performed by LC-MS/MS.

Sensitive, semi-quantitative Sensitive, quantitative High resolution In vivo or in vitro labeling No labeling necessary Bidifferential expression Small sample size Detects PTMs Sensitive, quantitative In vivo or in vitro labeling 4 stable isotope labels Detects PTMs Sensitive, quantitative Cell culture labeling Bidifferential expression Detects PTMs... 

Everley, P. A., Krijgsveld, J., Zetter, B. R., Gygi, S. P. (2004). Quantitative cancer proteomics stable isotope labeling with amino adds in cell culture (SILAC) as a tool for prostate cancer research. 

Pandey, A., Mann, M. (2002). Stable isotope labeling by amino adds in cell culture, SI LAC, as a simple and accurate approach to expression proteomics. 

SILAC Stable-isotope labeling with amino acids in cell culture... 

Labeling proteins with heavy and light tags and screening the hit compound versus an inactive control, followed by mass spectrometric comparison of the two samples, is another approach that avoids many of the common pitfalls in affinity methods (3). Techniques such as stable isotope labeling with amino acids in cell culture and isotope-coded affinity tagging (ICAT) exemplify these techniques. 

Quantification of Proteins with Stable-Isotope Labeling by Amino Acids in Cell Culture 469... 

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