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 (more than20mmol Ni/ml settled resin). The resin performs well in either column, batch or vacuum-based methods with a binding capacity of more than15mg/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 proteinsfrom 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.|
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.