SILAC Technique

1. Assignment on:
SILAC technique

3. SILAC
Technique
 Stable Isotope labeling by amino acids in cell culture

4. Introduction
 Stable isotope labeling using amino acids in cell culture (SILAC) is
a powerful method based on mass spectrometry that identifies
and quantifies relative differential changes in protein abundance.
 First used in quantitative proteomics in 2002, it provides accurate
relative quantification without any chemical derivatization or
manipulation.
 This technique detects differences in protein abundance among
sample using non-radioactive isotopic labeling.
 It is a popular method of quantitative proteomics.

5. principle  The principal of SILAC is based on metabolically incorporating
stable isotope labelled amino acids into the entire proteome.

7. Workflow Of SILAC

8. Applications
 Characterize protein quantitative differences between different
samples
 Investigate the changes of protein post translational modifications
 Distinguish specific interacting proteins in the protein-protein
interactions networks.

9. Advantages
and
limitation
 SILAC is simple and powerful method for quantitative analysis of
proteins characterized by quantitative accuracy and
reproducibility.
 Differentially treated samples can be combined at the level of
intact cells or proteins namely at the very first step of the
experimental workflow and can be processed together to
minimize experimental error.
 But it is only appropriate for cell samples, which requires a long
time due to cell culture.