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Products >  Protein_Expression_and_Purification >  His-Tagged_Protein_Purification >  Cobalt_Resin-FPLC

FPLC Purification of His-Tagged Proteins—TALON Superflow Metal Affinity Resin

TALON His-Tag Purification Resin lets you prepare exceptionally pure his-tagged proteins from bacterial, mammalian, yeast, and baculovirus-infected cells, under native or denaturing conditions. TALON is an immobilized metal affinity chromatography (IMAC) resin charged with cobalt, which binds to his-tagged proteins with higher specificity than nickel-charged resins. As a result, TALON resin delivers his-tagged proteins of the highest purity. In addition, each cobalt ion is bound to the resin at four sites, resulting in low metal ion leakage.

Reactive Core Contains Cobalt for Highest Purity

TALON Metal Affinity Resin is complexed with cobalt ions that make it highly selective for his-tagged proteins. TALON’s cobalt-containing reactive core has strict spatial requirements—only proteins containing adjacent histidines or specially positioned, neighboring histidines are able to bind. The spatial requirements for nickel-based resins (e.g., Ni-NTA) are less strict—these resins have a much higher affinity for randomly positioned (i.e., non-his-tag) histidines. As a result, TALON resin binds more specifically to his-tagged proteins.

TALON Superflow is specifically designed for quick and effective purification of his-tagged proteins at high flow rates and medium pressure (up to 150 psi)—it’s ideal for FPLC applications. For prepacked 1 ml and 5 ml cartridges, please see the HisTALON Cartridge product page.

Uniform Matrix Guarantees Less Contamination

Cobalt-based resins have a more uniform structure than nickel-based resins. TALON resin contains negatively charged reactive sites that form three-dimensional pockets. Each pocket contains three carboxyl groups and one nitrogen atom that collectively bind a single, positively charged cobalt ion—an arrangement that allows the cobalt ion to bind to two adjacent histidine residues. In this configuration, cobalt is bound very tightly and does not leach out of the resin. Nickel-based resins are less homogeneous in structure because nickel ions can form two different coordination complexes: one of which forms a three-dimensional pocket similar to that of the TALON ligand, and a second that forms a planar (flat) structure. In the distorted, planar structure, each nickel ion binds to only two carboxyl groups and one nitrogen atom. As a result, the planar structure binds the nickel ions less tightly, allowing them to leach from the resin. TALON Metal Affinity Resin, with its uniform matrix, provides high affinity binding under a variety of purification conditions, ensuring optimal protein purification.

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Features

  • Exhibits high affinity for his-tagged proteins
  • Ideal for FPLC applications
  • No copurification of proteins
  • Resists metal leakage
  • Performs well under a wide range of purification conditions

Applications

Purified recombinant his-tagged proteins can be used for:

  • Crystallography
  • Functional assays
  • Structural investigations
  • Other applications

 

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