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Home > Service > Protein Biomarker Discovery > The Choice Between NIT and SILAC labeling Techniques
The Choice Between NIT and SILAC labeling Techniques

The choice between NIT and SILAC labeling largely depends on the specific requirements of a project.

Both SILAC and NIT labeling can label virtually every peptides from digestion of a complex protein mixture. Both methods can be used to detect changes in protein abundance as well as changes in posttranslational modifications.

The major advantage of SILAC labeling is that the samples are metabolically labeled, so separations at protein level will not introduce error. So it is possible to fractionate samples at both protein level and peptide level, e.g. 2-D fractionation at protein level followed by 2-D fractionation at peptide level, although such a 4-dimentional fractionation will result a very large number of samples for MS analysis. The main disadvantage of SILAC method is that tissue or blood samples cannot be labeled by SILAC technique. Some cell lines are also difficult to grow in SILAC. The high cost of SILAC media is sometimes a concern too.

The major advantage of NIT method is its ability to label virtually any type of protein samples, including cells, tissue and blood samples. The cost of labeling reagents is very low compared to SILAC media. The main disadvantage is that labeling is performed after proteolytic digestion, so fractionation is carried out at peptide level. Normally it is limited to two-dimensional separations (SCX and RP-LC) before MS analysis.

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