Unloaded Immobilised Metal Affinity Chromatography (IMAC) Agaroses
Cambio offers high-performance immobilized metal affinity chromatography (IMAC) resins, based on BioWorks Workbeads in IDA or NTA formats. IDA or NTA agarose are supplied in unloaded form and can be loaded with transition metals to obtain different specificities for his-tagged proteins, or for other applications such as purification of phosphorylated proteins. If you require unloaded IMAC beads for purification from dilute starting material, then we recommend IDA or NTA MagBeads.
Which IMAC resin should I use to purify my protein (NTA or IDA?)
For immobilized-metal affinity chromatography (IMAC) you can choose between the metal ion being coupled to the resin matrix via either nitrilotriacetic acid (NTA) or iminodiacetic acid (IDA). There are other chelating ligands used for IMAC resins, but NTA and IDA are the popular ones. Which chelating ligand should you use and why? Does it make a difference?
NTA
- Tetravalent, coordination number 4
- Higher binding specificity
- Stronger coordination of metal ions
- Lower metal ion leaching
IDA
- Trivalent, coordination number 3
- Generally, more non-specific binding
- Increased metal ion leaching
- Higher metal loading density
- Requires lower eluent imidazole concentration
- Generally, lower yield
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