Application

HisTrap FF crude is a ready-to-use column, prepacked with precharged Ni Sepharose 6 Fast Flow. This prepacked column is intended for purification of histidine-tagged recombinant proteins by immobilized metal affinity chromatography (IMAC). After thorough cell disruption, it is possible to load the unclarified lysate on the column without precentrifugation and filtration of the sample. Extending the duration of the mechanical treatment of the sample to ensure a more complete lysis is recommended. For optimal results, we recommend first addition of lysozyme and DNase I followed by a mechanical lysis, for example by sonication. The homogenized sample can then be loaded directly on the column without a clarification step which may prevent degradation of the target protein and increase the activity.

Features and Benefits

Optimized for purification of histidine-tagged proteins directly from homogenized, unclarified cell lysates. High binding capacity, approx. 40 mg/ml medium and negligible Ni2+ leakage Compatible with a wide range of reducing agents, detergents, denaturants, and other additives. Reduced sample preparation time, which minimizes risk of protein degradation by proteases. No need for centrifugation or filtration of the samples. Direct sample application. Reliable purification of histidine-tagged proteins directly from unclarified lysates–simply sonicate and run. Reduced sample preparation time, which minimizes risk of protein degradation by proteases Simple operation with a syringe, pump, or chromatography system such as ÄKTAdesign or FPLC System Permit rapid yet reliable separations with a minimum of sample preparation and equipment

General description

HisTrap FF crude columns offer the convenience of simple, one-step purification of histidine-tagged proteins directly from homogenized, unclarified cell lysate by immobilized metal ion affinity chromatography (IMAC).

Legal Information

HisTrap is a trademark of Cytiva

Storage and Stability

Store at 4 to 30 °C (20% Ethanol)