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

In-vivo SILAC diets
In vitro SILAC is a well-established approach for quantification of proteins in cell culture. 1 The same concept
can also be applied to living organisms (in-vivo SILAC). The latter approach allows determination of protein
patterns of all tissues of an organism with respect to a reference organism. This approach has beensuccessfully applied to the analysis of mouse organs.2
Prerequisite for the application of the in-vivo SILAC approach is the availability of an appropriate SILAC dietin which all lysins are replaced by 13C-lysine.
Silantes provides SILAC diets for in-vivo labelling of mouse, but also for other model organisms such as fly 3
and worm4. To see our product portfolio of in-vivo SILAC diets, scan the QR-code in the top right corner.

Silantes SILAC Diets for in-vivo labelling of mice5
Mice are fed with Silantes SILAC mouse feed according to the scheme in Figure 2 on the next page forabout 90 days requiring about 800 g Silantes mice feed.
C-lysine labelled mice feed (Figure 1) has been developed in13

cooperation with the group of Prof. Matthias Mann, Max-PlanckInstitute of Biochemistry as a kit containing labelled and unlabelledmouse feed. The kit consists of 13C-lysine-labelled “heavy” diet (B)and unlabelled “light” diet (A). The feed is an artificial, amino acidbased feed using Harlan components.
After the metabolic labelling (feeding) of the mice according to thescheme in Figure 2, the mice are sacrificed. Differences in theFigure 1: Silantes 13C-lysine mice feed

protein patterns are determined in analogy to the established
SILAC approach in cell culture (see literature for Silantes in vitro SILAC). Instead of 13C-lysine labelled feed,other isotopic formulations are also available on request, e.g. 13C,15N-labelled feed.
1Ong, 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.
2Ong, S.E., and Mann, M. (2006). A practical recipe for stable isotope labelling by amino acids in cell culture (SILAC). Nat. Protoc. 1, 2650–2660.
3Sury, M. D., Chen, J. X., & Selbach, M. (2010). The SILAC fly allows for accurate protein quantification in vivo. Molecular & cellularproteomics : MCP, 9(10), 2173–2183. https://doi.org/10.1074/mcp.M110.000323
4

Larance, 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
5Krueger M., Moser M., Ussar S., Thievessen I., Luber Ch.A, Forner F., Schmidt S., Zanivan S., Faessler R. and Mann M.(2008). SILAC Mouse
for Quantitative Proteomics Uncovers Kindlin-3 as an Essential Factor for Red Blood Cell Function, Cell 134, 353–364.

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

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

SILAC spike-in using Silantes 13C6-Lysine-labelled mouse tissue
Figure 2 shows the SILAC spike-in workflow:
Differences in protein patterns of unlabelled
tissue (A) with respect to unlabelled tissue (B)can be quantified by “spiking-in” a 13C-labelledreference tissue (R). This 13C-lysine reference
tissue can be obtained from Silantes directly,
as frozen material or lyophilized (see literaturefor Silantes lyophilized tissue for SILAC spikein). To see our product portfolio of SILAC
mouse tissues, scan the QR-code in the topright corner.
Short outline of the procedure:
The isotopically labelled “heavy” reference
tissue (R) is mixed with the unlabelled “light”
tissues (A) and (B), respectively. The proteomesof the (A)(R)-mix and (B)(R)-mix are isolated,digested and subjected to LC-MS as shown in
the workflow. Therefore, the two peptide
amount ratios (A/R) and (B/R) can be

Figure 2:in-vivo SILAC-mouse workflow

determined.
Calculating the ratio (A/R) : (B/R) cancels out the reference amount (R) and yields the ratio of peptides (A/B).
The stable isotopically labelled mouse tissue (R) is used as a standard, permitting normalizing (A) withrespect to (B), accounting for differences in the isolation procedure in the mixtures (A/R) and (B/R) without
affecting the peptide ratios (A/B). (Strain (R) must not necessarily be the same as strain (A) and (B).)
Conclusion:

The customer is free in regard to the choice of strain used for theexperiment but can use the Silantes 13C6-Lysine-labelled mouse
tissue as a reference which is spiked into tissue (A) and (B).
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