| 货号 | 20432S |
| 供应商 | CST |
| 背景 | Immobilized metal affinity chromatography, or IMAC, has been widely used to enrich proteins and peptides from biological samples by binding to clusters of negative charge. Divalent transition metal ions Co2+, Cu2+, Ni2+, and Zn2+ are often used to purify proteins rich in poly-Histidine or Cysteine as well as proteins with metal affinity. Trivalent metal ions, Fe3+, Ga3+, Al3+, as well as Ti4+ and Zr4+ are commonly used for phospho-peptide enrichment for proteomic studies (1). Iminodiacetic acid (IDA) or nitrilotriacetic acid (NTA) are used for chelating the metal ions to agarose-coated beads. In comparison studies, NTA has been shown to perform better than IDA at selectively capturing and identifying more phospho-peptides. Ga3+ and Fe3+ are comparable with respect to the number of phospho-peptides identified (2). Compared to metal oxide affinity chromatography (MOAC) using TiO2, Fe3+ IMAC performed marginally better with TiO2 having a preference for acidic phospho-peptides (pI > 4) relative to Fe3+ which prefered less acidic peptides (pI < 4) (3). The PTMScan® Fe-NTA Magnetic Beads offer an efficient tool for phospho-peptide enrichment with little or no bias for phospho-residue context. They can be employed independently or in conjunction with immunoaffinity based enrichment to complement any PhosphoScan® LC-MS/MS proteomic study. |
| 运输条件 | 0.75 |
| 存放说明 | 4C |
| 参考文献 | 1 . Block, H. et al. (2009) Methods Enzymol 463, 439-73. 2 . Ficarro, S.B. et al. (2009) Anal Chem 81, 4566-75. 3 . Yue, X. et al. (2015) Anal Chem 87, 8837-44. |
This chart shows the underlying motif distribution in a PhosphoScan® IMAC LC-MS/MS experiment using 4026 nonredundant tryptic peptides generated from mouse embryo and captured using PTMScan® Phospho-Enrichment IMAC Fe-NTA Magnetic Beads. Within this data set, 84% of the peptides were phosphorylated on serine residues, 13% on threonine, and 3% on tyrosine. | |
The Motif Logo was generated from a PhosphoScan® IMAC LC-MS/MS experiment using 4026 nonredundant tryptic peptides derived from mouse embryo and captured using PTMScan® Phospho-Enrichment IMAC Fe-NTA Magnetic Beads. The logo demonstrates the relative frequency of amino acids in a given position around the modified site within this data set. |
