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HisTrap™ FF crude Columns/HisTrap FF crude Kit - Convenient Purification from Unclarified Lysates

• Optimized for purification of histidine-tagged* proteins directly from homogenized, unclarified cell lysates.
• Simply homogenize and load your sample directly.
• 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.
• Convenient kit provides columns and buffers for quick and easy purifications.
• Simple operation with a syringe, pump, or chromatographic system such as ÄKTAdesign.

HisTrap™ FF crude Columns/HisTrap FF crude Kit - Convenient Purification from Unclarified Lysates

Technical Information

* For licensing information, see Legal Info.

Customer feedback on Ni Sepharose 6 Fast Flow.

TECHNICAL SPECIFICATIONS
Medium	Ni Sepharose 6 Fast Flow
Column volume	1 ml and 5 ml
Column dimensions	0.7 × 2.5 cm (1 ml) 1.6 × 2.5 cm (5 ml)
Dynamic binding capacity*	Approx. 40-mg histidine-tagged protein/ml medium
Recommended flow rate†	1 ml/min (1 ml) 5 ml/min (5 ml)
Max. pressure†	3 bar (0.3 MPa, 42 psi)
Compatibility  during use	Stable in all commonly used buffers, reducing agents, denaturing agents and detergents, see Ni Sepharose™ High Performance - High-performance Purification
Chemical stability‡	For one week at 40°C: 0.01 M HCl, 0.1 M NaOH For 12 h: 1 M NaOH, 70% acetic acid 30 min tested: 30% 2-propanol 1 h tested: 2% SDS
pH stability‡	2-14 (short term), 3-12 (long term)
Storage	20% ethanol
Storage temperature	+4°C to 30°C
* Protein-dependent † H2O at room temperature ‡ Ni2+-stripped medium

HisTrap™ FF crude columns allow simple, one-step purification of histidine-tagged proteins without sample pretreatment such as centrifugation and filtration.

HisTrap™ FF crude 1-ml and 5-ml columns offer the convenience of simple, one-step purification of histidine-tagged proteins directly from homogenized, unclarified cell lysates. Direct loading of unclarified sample both saves time and prevents degradation of sensitive target protein.

HisTrap™ FF crude columns are prepacked with Ni Sepharose™ 6 Fast Flow. For more information see Ni Sepharose™ 6 Fast Flow - Scale-up Purification. For column technical specifications, see HisTrap™ FF Columns - Easy Scale-up Purification (HisTrap™ FF Columns). For best results, we recommend first addition of lysozyme and DNase I followed by a mechanical lysis, e.g. 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.

HisTrap FF crude Kit provides convenient purification of histidine-tagged proteins. The kit contains three HisTrap FF crude 1-ml columns, phosphate buffer, pH 7.4 (2 × 50 ml [8× concentrate]), 2 M imidazole, pH 7.4, all necessary connectors, a syringe, and instructions.

Four repeated purifications of MBP-(His)₆ from E. coli extract on HisTrap FF crude 1 ml. The medium was subjected to Ni²⁺-ion stripping, cleaning-in-place, and recharging between runs.

HisTrap FF crude columns save time in the purification of histidine-tagged proteins compared to conventional IMAC.