Isotope-coded affinity tagging

In vitro labeling of proteins using isotope-coded affinity tags... 

Gygi, S. P., Rist, B., Gerber, S. A., Turecek, F., Gelb, M. H., and Aebersold, R. (1999). Quantitative analysis of complex protein mixtures using isotope-coded affinity tags. Nat. Biotechnol. 17, 994-999. 

Von Haller, P.D., Yi, E., Donohoe, S., Vaughn, K., Keller, A., Nesvizhskii, A.I., Eng, J., Li, X.J., Goodlett, D.R., Aebersold, R., Watts, J.D. (2003). The Application of New Software Tools to Quantitative Protein Profiling Via Isotope-coded Affinity Tag (ICAT) and Tandem Mass Spectrometry II. Evaluation of Tandem Mass Spectrometry Methodologies for Large-Scale Protein Analysis, and the Application of Statistical Tools for Data Analysis and Interpretation. Mol. Cell. Proteomics 2, 428 -42. 

Fauq, A.H., Kache, R., Khan, M.A., and Vega, I.E. (2006) Synthesis of acid-cleavable light isotope-coded affinity tags (ICAT-L) for potential use in proteomic expression profiling analysis. Bioconjugate Cbem. 17, 248-254. 

Han, B.N., Stevens, J.F., and Maier, C.S. (2007) Design, synthesis, and application of a hydrazide-func-tionalized isotope-coded affinity tag for the quantification of oxylipid-protein conjugates. Anal. Chem. 79(9), 3342-3354. 

Hansen, K.C., Schmitt-Ulms, G., Chalkley, R.J., Hirsch, J., Baldwin, M.A., and Burlingame, A.L. (2003) Mass spectrometric analysis of protein mixtures at low levels using cleavable 13C-isotope-coded affinity tag and multidimensional chromatography. Mol. Cell. Proteomics 2, 299-314. 

Li, J., Steen, H., and Gygi, S.P. (2003) Protein profiling with cleavable isotope-coded affinity tag (cICAT) reagents. The yeast salinity stress response. Mol. Cell. Proteomics 2, 1198-1204. 

Lu, Y., Bottari, P., Turecek, F., Aebersold, R., and Gelb, M.H. (2004) Absolute quantification of specific proteins in complex mixtures using visible isotope-coded affinity tags. Anal. Chem. 76, 4104 flll. 

Turecek, F. (2002) Mass spectrometry in coupling with affinity capture-release and isotope-coded affinity tags for quantitative protein analysis. J. Mass Spectrom. 37, 1-14. 

The isotope-coded affinity tag approach utilizes chemical labeling that allows quantitation when combined with mass spectrometry. ICAT is desirable because the chemical labeling takes advantage of the mass defects of monoisotopic stable isotopes. ICAT uses an ICAT reagent to differentially label protein samples on their cysteine residues. ICAT is advantageous because it permits the evaluation of low-abundance proteins and proteins at both extremes of molecular weights and isoelectric points.60... 

S. P. Gygi, B. Rist, S. A. Gerber, F. Turecek, M. H. Gelb, and R. Aebersold. Quantitative Analysis of Complex Protein Mixtures Using Isotope-Coded Affinity Tags. Nat. Biotechnol., 17(1999) 994-999. 

D.K. Han, J. Eng, H. Zhou and R. Aebersold, Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags (ICAT) and mass spectrometry, Nat. Biotech., 19 (2001) 946-951. 

Mass spectrometry — It has excellent sensitivity but low throughput and is semiquantitative. The ICAT (isotope-coded affinity tag) labeling method for proteins enables differential display analyses using mass spectrometry (Griffin and Abersold, 2001). This may be... 

Gygi SP, Rist B, Gerber SA et al. Quantitative analysis of complex protein mixtures using isotope-coded affinity tags. NatBiotechnol. 1999 17 994-999. 

Li C, Hong Y, Tan YX et al. Accurate qualitative and quantitative proteomic analysis of clinical hepatocellular carcinoma using laser capture microdissection coupled with isotope-coded affinity tag and two-dimensional liquid chromatography mass spectrometry. Mo/ Cell Proteomics 2004,3399-409. 

Affinity capture-release electrospray ionization mass spectrometry (ACESIMS) and isotope-coded affinity tags (ICAT) are two recently introduced techniques for the quantitation of protein activity and content with applications to clinical enzymology... 

More importantly insoluble proteins, such as membrane-bound and nuclear proteins, are under-represented within 2DE analysis. However, membrane proteins can be identified by MS following a IDE separation. Unfortunately, hydrophobicity is the most common reason for poor representation of abundant proteins and seems set to be with us for quite some time (Santoni et al., 2000 Rabilloud, 2002). Emerging proteomic methods without the use of 2DE are being developed, such as isotope-coded affinity tagging (ICAT) and multi-dimensional protein identification technology (MuDPIT). 

Paba, J., Ricart, C.A.O., Fontes, W., Santana, J.M., Teixeira, A.R.L., Marchese, J., Williamson, B., Hunt, T., Karger, B.L. and Sousa, M.V. (2004) Proteomic analysis of Trypanosoma cruzi developmental stages using isotope-coded affinity tag reagents. Journal of Proteome Research 3, 51 7-524. 

Smolka MB, Zhou H, Purkayastha S, Aebersold R. 2001. Optimization of the isotope-coded affinity tag-labeling procedure for quantitative proteome analysis. Anal Biochem 297 25-31. 

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