HisLink™ Protein Purification Systems

HisLink™ Protein Purification Systems are designed for purification of polyhistidine (His)-tagged or HQ-tagged proteins directly from culture medium containing bacterial cells expressing the tagged protein of interest. All product formats use the HisLink™ Protein Purification Resin (Cat.# V8821), a macroporous silica resin derivatized with a high level of tetradentate-chelated nickel (>20mmol Ni/ml settled resin). The resin performs well in either column, batch or vacuum-based methods with a binding capacity of >15mg/ml of resin. The HisLink™ Protein Purification Resin is useful in all general ...

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HisLink™ Protein Purification Systems are designed for purification of polyhistidine (His)-tagged or HQ-tagged proteins directly from culture medium containing bacterial cells expressing the tagged protein of interest. All product formats use the HisLink™ Protein Purification Resin (Cat.# V8821), a macroporous silica resin derivatized with a high level of tetradentate-chelated nickel (>20mmol Ni/ml settled resin). The resin performs well in either column, batch or vacuum-based methods with a binding capacity of >15mg/ml of resin. The HisLink™ Protein Purification Resin is useful in all general immobilized metal affinity chromatography (IMAC) applications matrix as well as in low- to medium-pressure liquid chromatography systems.

The bacterial cells are lysed using the FastBreak™ Cell Lysis Reagent (Cat.# V8573), and the crude lysate is combined with the HisLink™ Resin. The addition of these reagents results in simultaneous bacterial lysis and binding of the polyhistidine- or HQ-tagged proteins. The samples are transferred to user-provided columns or to included Spin Columns (Cat.# V1320), where the untagged proteins are washed away and the His-tagged protein is recovered by elution with imidazole. If desired, the resin may be used in vacuum filtration devices (e.g., Vac-Man® Vacuum Manifold [Cat.# A7231]) to rapidly process simultaneous samples. The HisLink™ 96 Protein Purification System (Cat.# V3680) provides a simple and quick vacuum-based method of simultaneously purifying multiple His-tagged proteins  from E. coli cultured in deep-well, 96-well plates. The HisLink™ 96 System is amenable to manual or automated methods, such as the Beckman Coulter Biomek® 2000 or FX for high-throughput applications, yielding 1mg of purified polyhistidine-tagged protein per well.

Features - Benefits

• Save Time: No centrifugation (pre-clearing) required; polyhistidine- or HQ-tagged proteins are purified directly from cleared or crude cell lysates.
• Quick: No long lysozyme incubations are required for cell lysis.
• Flexible and Versatile: Perform purification manually in batch, using a vacuum manifold, using liquid chromatography or liquid handling platform.

Applications

• Protein purification of polyhistidine- or HQ-tagged proteins using IMAC.
• Purification of tagged proteins from E. coli using a vacuum- or centrifugation-based format.
• Protein purification of polyhistidine- or HQ-tagged proteins from 96-well, deep-well culture plates using IMAC.

For more information, see the Protocols & Applications Guide.

Notes

Resin product volumes represent total volume of a 50% resin slurry.

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