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SILAC (Stable isotope labelling with amino acids in cell culture) is a mass spectrometry
Mass spectrometry

Mass spectrometry is an analytical technique for the determination of the elemental composition of a sample or molecule. It is also used for elucidating the chemical structures of molecules, such as peptides and other chemical compounds....
-based technique developed to detect differences in protein abundance between two (or more) samples (Ong et al., 2002). It is one of the most popular methods for quantitative proteomics
Quantitative proteomics

The aim of quantitative proteomics is to obtain quantitative information about all proteins in a sample. Rather than just providing lists of proteins identified in a certain sample, quantitative proteomics yields information about differences between samples....
. Two populations of cells are cultivated in cell culture
Cell culture

Cell culture is the process by which prokaryote or eukaryote cells are grown under controlled conditions. In practice the term "cell culture" has come to refer to the culturing of cells derived from multicellular eukaryotes, especially animal cells....
. One of the cell populations is fed with growth medium
Growth medium

A growth medium or culture medium is a liquid or gel designed to support the growth of microorganisms or Cell s , or small plants like the moss Physcomitrella patens ....
 containing normal amino acids. In contrast, the growth medium of the second cell population contains amino acids labeled with stable (non-radioactive) heavy isotope
Isotope

Isotopes are any of the different types of atoms of the same chemical element, each having a different atomic mass . Isotopes of an element have atomic nucleus with the same number of protons but different numbers of neutron....
s.






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SILAC (Stable isotope labelling with amino acids in cell culture) is a mass spectrometry
Mass spectrometry

Mass spectrometry is an analytical technique for the determination of the elemental composition of a sample or molecule. It is also used for elucidating the chemical structures of molecules, such as peptides and other chemical compounds....
-based technique developed to detect differences in protein abundance between two (or more) samples (Ong et al., 2002). It is one of the most popular methods for quantitative proteomics
Quantitative proteomics

The aim of quantitative proteomics is to obtain quantitative information about all proteins in a sample. Rather than just providing lists of proteins identified in a certain sample, quantitative proteomics yields information about differences between samples....
. Two populations of cells are cultivated in cell culture
Cell culture

Cell culture is the process by which prokaryote or eukaryote cells are grown under controlled conditions. In practice the term "cell culture" has come to refer to the culturing of cells derived from multicellular eukaryotes, especially animal cells....
. One of the cell populations is fed with growth medium
Growth medium

A growth medium or culture medium is a liquid or gel designed to support the growth of microorganisms or Cell s , or small plants like the moss Physcomitrella patens ....
 containing normal amino acids. In contrast, the growth medium of the second cell population contains amino acids labeled with stable (non-radioactive) heavy isotope
Isotope

Isotopes are any of the different types of atoms of the same chemical element, each having a different atomic mass . Isotopes of an element have atomic nucleus with the same number of protons but different numbers of neutron....
s. For example, the medium can contain arginine
Arginine

Arginine is an a-amino acid. The Optical isomerism is one of the 20 most common natural amino acids. Its codons are CGU, CGC, CGA, CGG, AGA, and AGG....
 labeled with six carbon-13
Carbon-13

Carbon-13 is a natural, Stable isotope isotope of carbon and one of the environmental isotopes. It makes up about 1.1% of all natural carbon on Earth....
 atoms (13C) instead of the normal carbon-12
Carbon-12

Carbon-12 is the most Abundance of the two Stable_isotope isotopes of the element carbon, accounting for 98.89% of carbon; it contains 6 protons, 6 neutrons, and 6 electrons....
 (12C). When the cells are growing in this medium, they incorporate the heavy arginine into all of their proteins. Therefore, all of the arginine containing peptides are now 6 Da heavier than their normal counterparts. The trick is that the proteins from both cell populations can be combined and analyzed together by mass spectrometry
Mass spectrometry

Mass spectrometry is an analytical technique for the determination of the elemental composition of a sample or molecule. It is also used for elucidating the chemical structures of molecules, such as peptides and other chemical compounds....
. Pairs of chemically identical peptides of different stable-isotope composition can be differentiated in a mass spectrometer owing to their mass difference. The ratio of peak intensities in the mass spectrum for such peptide pairs accurately reflects the abundance ratio for the two proteins. SILAC has emerged as a very powerful method to study cell signaling
Cell signaling

Cell signaling is part of a complex system of communication that governs basic cellular activities and coordinates cell actions. The ability of cells to perceive and correctly respond to their microenvironment is the basis of development, tissue repair, and immunity as well as normal tissue homeostasis....
, protein-protein interaction
Protein-protein interaction

Protein-protein interactions involve the association of protein molecules. These associations are studied from the perspective of biochemistry, signal transduction and graph theory....
 and regulation of gene expression
Regulation of gene expression

Gene modulation redirects here. For information on therapeutic regulation of gene expression, see therapeutic gene modulation.Regulation of gene expression includes the processes that cell s and viruses use to turn the information on genes into gene products....
.

See also

  • Cell signaling
    Cell signaling

    Cell signaling is part of a complex system of communication that governs basic cellular activities and coordinates cell actions. The ability of cells to perceive and correctly respond to their microenvironment is the basis of development, tissue repair, and immunity as well as normal tissue homeostasis....
  • mass spectrometry
    Mass spectrometry

    Mass spectrometry is an analytical technique for the determination of the elemental composition of a sample or molecule. It is also used for elucidating the chemical structures of molecules, such as peptides and other chemical compounds....
  • Protein-protein interaction
    Protein-protein interaction

    Protein-protein interactions involve the association of protein molecules. These associations are studied from the perspective of biochemistry, signal transduction and graph theory....
  • Proteomics
    Proteomics

    Proteomics is the large-scale study of proteins, particularly their protein structure and functional genomics. Proteins are vital parts of living organisms, as they are the main components of the physiological metabolic pathways of biological cell....
  • Quantitative proteomics
    Quantitative proteomics

    The aim of quantitative proteomics is to obtain quantitative information about all proteins in a sample. Rather than just providing lists of proteins identified in a certain sample, quantitative proteomics yields information about differences between samples....
    • ICAT
      Isotope-coded affinity tag

      Isotope-coded affinity tags is a gel-free method for quantitative proteomics that relies on chemical labeling reagents referred to as ICATs. These chemical probes consist of 3 general elements: a reactive group capable of labeling a defined amino acid side chain , an isotopically coded linker, and a tag for the affinity isolation of labeled...
    • Tandem mass tags
      Tandem mass tags

      Tandem Mass Tags are chemical labels used for Mass_spectrometry based quantitation and identification of biological macromolecule such as Protein, Peptides and Nucleic_acids....
    • iTRAQ
      ITRAQ

      iTRAQ is a non-gel based technique used to identify and quantify proteins from different sources in one single experiment. It uses isotope coded covalent tags....
    • Label Free Quantitation


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