Thermo Scientific HisPur Cobalt Superflow Agarose is a tetradentate-chelating Superflow 6 support charged with divalent cobalt, designed for high-purity, large-scale purification of polyhistidine-tagged proteins.
Features of HisPur Cobalt Superflow Agarose:
• Robust—highly crosslinked 6% agarose Superflow beaded support tolerates linear flow rates up to 1200 cm/hour with minimal leaching of cobalt due to a tetradentate chelator • High yields—binds 20 mg of 6xHis-GFP per mL of resin at a linear flow rate of 150 cm/hour, can bind greater than 30 mg per mL of resin depending on flow rate • High purity—specificity of cobalt binding to histidine-tag generally results in elution fractions with less than 10% contamination of non-specific proteins • Compatible—maintains function after exposure to a wide variety of chemicals and pH values • Cost effective—resin is stable through multiple cycles of cleaning and reuse
HisPur Cobalt Superflow Agarose is a highly crosslinked, durable resin that does not compress at flow rates used for medium- to large-scale FPLC purifications. The resin holds up well to a variety of chemicals and pH values, and is compatible with common clean-in-place procedures. Compared to nickel and other ligands commonly used for immobilized metal affinity chromatography (IMAC), cobalt provides greater specificity for His-tagged protein purification. The specific binding of HisPur Cobalt Superflow Agarose results in purer elution fractions, reducing the need for extra polishing steps.
Applications: • Large scale FPLC purification of polyhistidine tagged proteins
Affinity chromatography is often used as a quick and easy approach for the purification of recombinant proteins. One such method of purification involves the binding of a polyhistidine tag to a divalent metal cation that has been coordinated by a chelator immobilized on a beaded support. For immobilized metal affinity chromatography (IMAC) purification of His-tagged proteins, the type of bead, chelator, and metal immobilized influences purity, yield, and flow rate performance of these purifications.
The highly crosslinked 6% agarose Superflow beaded support tolerates larger linear flow rates than agarose resins that have lower amounts of crosslinking. The HisPur Cobalt Superflow Agarose protocol utilizes gentle wash and elution conditions and typically produces greater than 90% pure target protein. Protein purities achieved with HisPur Cobalt Superflow Agarose are generally higher than those achieved with nickel IMAC resins making HisPur Cobalt Superflow Agarose a valuable tool for users interested in one-step purifications. Additionally, HisPur Cobalt Superflow Agarose is perfect for larger scale purifications that require the resin to withstand medium pressures without compressing.