Description
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).
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.
Ni Sepharose™ 6 Fast Flow has low nickel ion (Ni²⁺) leakage and is compatible with a wide range of additives used in protein purification. The special design of the column in combination with the medium, provide fast, simple, and convenient purifications. Short purification time generally minimizes deleterious effects, such as degradation and oxidation of sensitive target proteins, and is therefore of great importance. HisTrap™ FF crude columns can be operated with a syringe, peristaltic pump, or liquid chromatography system such as AKTA design chromatography systems.
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 AKTAdesign or FPLC™ System
Permit rapid yet reliable separations with a minimum of sample preparation and equipment
Preparation Note
Please be aware this product may be shipped 90 days before the expiration date. For more information on the batch specific expiration date, please contact technical service.
Store at 4 to 30 °C (20% Ethanol)
Analysis Note
To view the Certificate of Analysis for this product, please visit www.cytiva.com.
Other Notes
1)The pressure over the packed bed varies depending on a range of parameters such as the characteristics of the chromatography medium and the column tubing used.
The pressure over the packed bed varies depending on a range of parameters such as the characteristics of the chromatography medium and the column tubing used.
Legal Information
FPLC is a trademark of Cytiva
HisTrap is a trademark of Cytiva
Sepharose is a trademark of Cytiva
