Immobilized metal affinity chromatography, or IMAC, has been employed for many applications making use of its versatile interaction with aromatic nitrogens and phosphates. To this end, NTA matrices have been loaded with a broad range of transition metals, as optimized for the individual interaction. Magnetic beads are ideal for protein purification from dilute supernatants and for pull-down experiments. Al-NTA MagBeads are often used for the purification of phosphorylated proteins. Alternative matrices are Fe-, Ga- or Zr-NTA. All PureCube IMAC MagBeads are delivered as a 25% suspension.
PureCube Al-NTA MagBeads provide:
- Ready-to-use magnetic beads
- Purification of phosphorylated proteins
- Al-NTA Agaroses also available
Purification of phosphorylated proteins
IMAC methods, in particular Al-IDA, Fe-NTA or Ga-NTA, have been widely used to enrich phosphoproteins and phosphopeptides as part of the sample preparation for mass spectrometry (1,3,4). Depending on the sample, and the kind of phosphorylated proteins to be analyzed, also other transition metals, such as zirconium or aluminium have been loaded on NTA or IDA matrices for enrichment (5). Also, magnetic beads have been useful for this kind of sample preparation (2).
||Specific binding of phosphorylated proteins
||Affinity to phosphorylated proteins
||Depending on the application
||Delivered as a 25 % suspension
|Metal Ion capacity
||>12 µeqv Al3+/mL.
1. Albuquerque, C.P. et al. A multidimensional chromatography technology for in-depth phosphoproteome analysis. Mol Cell Proteomics (2008), 7(7), 1389-1396.
2. Herskowitz, J., et al. Phosphoproteomic analysis reveals site-specific changes in GFAP and NDGR2 phosphorylation in frontotemporal lobar degeneration. J. Proteome Res (2010), 9(12):6368-6379.
3. Yu, P. et al. Global analysis of neuronal phosphoproteome regulation by chondroitin sulfate proteoglycans. PLoS One (2013), 8,3, e59285.
4. Aryal, U.K. et al. Optimization of immobilized Gallium (III) ion affinity chromatography for selective binding and recovery of phosphopeptides from protein digests. Journal of Biomolecular Techniques (2008), 19:296-310.
5. Block et. al. Immobilized-metal affinity chromatography (IMAC) a review. Methods Enzymol. (2009), 463:439-73.