Final answer:
IMAC offers high selectivity for phosphopeptides, especially useful in phosphoproteomics. It can be combined with other techniques for improved quantification and offers a single-step purification process, ensuring higher reproducibility and robust analysis.
Step-by-step explanation:
One of the advantages of using Immobilized Metal Affinity Chromatography (IMAC) for protein purification over more traditional forms of chromatography includes its high selectively for phosphopeptides and phosphoproteins, which are often present in low abundances. This makes IMAC particularly useful in studies of phosphoproteomics, where the identification and characterization of phosphorylation sites are crucial, as seen in the analysis of the Arabidopsis plasma membrane. Additionally, IMAC can be combined with other techniques such as high-pH reverse phase separation for in-depth proteomic profiling, enhancing the recovery of phosphoproteins and reducing the complication of analyzing complex protein mixtures.
The combined use of IMAC with other labeling techniques, like isobaric Tags for Relative and Absolute Quantitation (iTRAQ) and Stable Isotope Labeling by Amino acids in Cell culture (SILAC), allows for the improved quantification of phosphorylation profiles. Another advantage is that IMAC enables the purification of phosphoproteins in a single step, which is beneficial in preserving the phosphorylation state that might be affected by multi-step processes. Moreover, IMAC tends to show high technical reproducibility and robustness, as demonstrated by comparisons with other methods such as Metal Oxide Affinity Chromatography (MOAC).