Scan the QR-Code to see our productcatalogue of SILAM Mouse Diets.

In-vivo SILAM diet
Stable isotopic labelling is the most reliable and accurate method for quantitative proteomics andmetabolomics. Proteomics studies on uniformly 15N-labelled rats have been previously reported using
N-labelled algae as protein source for Stable Isotope Metabolic Labelling in Mammals (SILAM).1

15

Silantes provides SILAM diets for in-vivo labelling of mouse, but also for other model organisms such as
fly2 and worm3 (see literature in the footnote). To see our product portfolio of in-vivo SILAM diets, scan theQR-code in the top right corner.

Silantes 15N-SILAM Diets for in-vivo labelling of mice
Silantes has developed a 15N-diet in cooperation with thegroup of Professor Chris Turck, Max-Planck-Institute of
Psychiatry as a kit containing a 15N-labelled “heavy” diet (B)and unlabelled “light” diet (A).4 It is an artificial feed using
Harlan components and a hydrolysate of the bacteria

Ralstonia

eutropha

asN-protein

15source.

Feedingexperiments showed that the mice accepted bacterial-based
protein better than algae-based protein.

Figure 1: Silantes 15N-labelled mice feed

After the metabolic labelling (feeding)of the mice according to the scheme infigure 2 or figure 3, the mice aresacrificed. Differences in the protein
patterns are determined in analogy tothe established SILAC approach in cell
culture (see literature for Silantes in vitro
SILAC).
Figure 2: Partial in-vivo labelling for global protein turnover

1

Daniel B. McClatchy, Meng-Qiu Dong, Christine C. Wu, John D. Venable, and John R. Yates (2007). 15N Metabolic Labeling of Mammalian
Tissue with Slow Protein, Journal of Proteome Research, 6(5), 2005-2010.
2Sury, M. D., Chen, J. X., & Selbach, M. (2010). The SILAC fly allows for accurate protein quantification in vivo. Molecular & cellular
proteomics : MCP, 9(10), 2173–2183. https://doi.org/10.1074/mcp.M110.000323
3Larance, M., Bailly, A. P., Pourkarimi, E., Hay, R. T., Buchanan, G., Coulthurst, S., Xirodimas, D. P., Gartner, A., & Lamond, A. I. (2011).
Stable-isotope labeling with amino acids in nematodes. Nature methods, 8(10), 849–851. https://doi.org/10.1038/nmeth.1679
4Zhang, M.D. Filiou, G. Chen, H. Heumann, Chris W. Turck (2008). Biomarker discovery in a mouse model of trait anxiety using 15N
metabolic labeling, Poster HUPO Amsterdam.

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Scan the QR-Code to see our productcatalogue of SILAM Mouse Tissues.

In-vivo SILAM tissue
A variation of the in-vivo SILAM-approach is the “spike-in” approach.

SILAM spike-in using Silantes 15N-labelled mouse tissue
Figure 2 shows the SILAM spike-in workflow:
N Reference

Differences in protein patterns of unlabelled case

N Control

15

N Case1414

tissue with respect to unlabelled control tissue canbe quantified by “spiking-in” aN-labelled15reference tissue.
This 15N reference tissue can be obtained from
Silantes directly, as frozen material or lyophilized(see literature for Silantes lyophilized tissue for
SILAM spike-in).
N Case14

N Reference

N Control

15

14

To see our product portfolio of SILAM mouse

N Reference

15

tissues, scan the QR-code in the top right corner.
N Case14

N Control

14Figure 3:Uniform in-vivo labelling of SILAM mouse

Short outline of the procedure:
The isotopically labelled “heavy” reference tissue is mixed with the unlabelled “light” case and control tissue,respectively. The proteomes of the case/reference-mix and control/reference-mix are isolated, digested
and subjected to LC-MS. Therefore, the two peptide amount ratios can be determined in analogy to thein-vivo SILAC-workflow to determine the case/control ratio (see Silantes in-vivo SILAC literature).
In addition to spiking-in the 15N-tissue as reference material for relative protein quantification by mass
spectrometry, the SILAM approach is used in further applications such as-Partially labelled mouse specimens for global protein turnover analysis by mass spectrometry (seefigure 2)5-Labelled metabolite standards for molecular structure confirmation-Biomarker discovery and verification

5Zhang, Reckow S, Webhofer C, Boehme M, Gormanns P, Egge-Jacobsen WM, Turck CW. (2011) Proteome scale turnover analysis in live
animals using stable isotope metabolic labeling. Anal Chem 83:1665-1672.

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