![]() Nickel or cobalt metals immobilized by NTA-chelation chemistry are the systems of choice for this application (see next section). The particular metal and chelation chemistry of a support determine its binding properties and suitability for specific applications of IMAC.Īffinity purification of His-tagged fusion proteins is the most common application for metal-chelate supports in protein biology research. Using nickel as the example metal, the resulting affinity support is usually called Ni-chelate, Ni-IDA or Ni-NTA resin. ![]() Once IDA-agarose or NTA-agarose resin is prepared, it can be "loaded" with the desired divalent metal (e.g., Ni, Co, Cu, and Fe). The chelators most commonly used as ligands for IMAC are nitrilotriacetic acid (NTA) and iminodiacetic acid (IDA). IMAC is a widely-used method for rapidly purifying polyhistidine affinity-tagged proteins, resulting in 100-fold enrichments in a single purification step. This basis for affinity purification is known as immobilized metal affinity chromatography (IMAC). Supports such as beaded agarose or magnetic particles can be derivatized with chelating groups to immobilize the desired metal ions, which then function as ligands for binding and purification of biomolecules of interest.
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