Skip to main content

Stable-Isotope Labeling for Protein Quantitation by Mass Spectrometry

Buy Article:

$55.00 plus tax (Refund Policy)

Mass spectrometry has become a routine instrument to identify proteins and peptides from simple or complex samples. Although identification can be confidently determined from a single experiment, quantitation requires multiple replicates and careful analysis. Alternatively, stable isotopes can be used to obtain relative quantitation of proteins and peptides from fewer replicates. Conventionally, half of a sample is labeled with stable isotope and mixed with the other half of unlabeled sample. The mixed sample is analyzed by mass spectrometry and because the stable isotope does not change the chemical properties of the peptide, the intensities of the unlabeled and labeled peptide can be directly compared. Absolute quantitation is obtained by adding a known amount of stable isotope labeled peptide or protein and comparing to an unlabeled counterpart. Stable isotope labeling methodologies can be divided into three categories: Chemical, enzymatic and metabolic. Here we provide an up-to-date review comparing the benefits and drawbacks of all three stable isotope labeling methodologies and briefly describe quantitation software solutions. In addition to quantitation, stable isotopes have also been used to identify post-translational modifications in proteins, identify components of DNA-protein and protein-protein complexes and to distinguish background contaminants from experimental results. Finally, we describe how fragmentation patterns from stable isotope labeled peptide and unlabeled peptides can improve peptide and protein identification and validation.





No References
No Citations
No Supplementary Data
No Data/Media
No Metrics

Keywords: ICAT; Mass spectrometry; SILAC; chemical; enzymatic; iTRAQ; isobaric; labeling; metabolic; proteomics; quantitation; software; stable isotope

Document Type: Research Article

Publication date: 2010-07-01

More about this publication?
  • Current Proteomics research in the emerging field of proteomics is growing at an extremely rapid rate. The principal aim of Current Proteomics is to publish well-timed review articles in this fast-expanding area on topics relevant and significant to the development of proteomics. Current Proteomics is an essential journal for everyone involved in proteomics and related fields in both academia and industry.
  • Access Key
  • Free content
  • Partial Free content
  • New content
  • Open access content
  • Partial Open access content
  • Subscribed content
  • Partial Subscribed content
  • Free trial content
Cookie Policy
X
Cookie Policy
Ingenta Connect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more