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. Both the "spin" and "96" 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. 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. No long lysozyme incubations are required for cell lysis.
The resin can be used with 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.
Note: Resin product volumes represent total volume of a 50% resin slurry.
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.