Ni Sepharose™ 6 Fast Flow - Scale-up Purification

• Designed for scaling up purification of histidine-tagged* proteins.
• Suitable for gravity-flow purification and multiwell plate screening.
• High protein binding capacity.
• Negligible leakage of Ni²⁺ ions.
• Compatible with a wide range of reducing agents, detergents, and other additives

Ni Sepharose™ 6 Fast Flow consists of 90-μm beads of highly cross-linked agarose, to which a chelating ligand has been immobilized. This chelating ligand is charged with Ni²⁺ ions, the first-choice metal ion for purifying most histidine-tagged protein. The ligand density of Ni Sepharose 6 Fast Flow ensures high binding capacity while leakage of Ni²⁺ ions is negligible, which ensures reliable capture of histidine-tagged proteins in repeated IMAC purifications. The high flow rate property of the Sepharose 6 Fast Flow matrix makes it ideal for scaling up purposes.

Ni Sepharose 6 Fast Flow is designed for scaling up purification of histidine-tagged proteins, gravity-flow purification, and multiwell plate screening.

Ni Sepharose™ 6 Fast Flow - Scale-up Purification

Technical Information

For more information on these products and the complete range of products for purification of histidine-tagged proteins, see Introduction to Tagged Protein Purification.

Ni Sepharose 6 Fast Flow is supplied preswollen in 20% ethanol. The medium is also available prepacked in 1-ml and 5-ml HisTrap FF and 20-ml HisPrep FF 16/10 columns, see HisPrep™ FF 16/10 Column - Easy Scale-up Purification, 1-ml and 5-ml HisTrap FF crude columns, see HisTrap™ FF crude Columns/HisTrap FF crude Kit - Convenient Purification from Unclarified Lysates, His MultiTrap FF 96-well plates, see His MultiTrap 96-Well Filter Plates - High-throughput Screening, and in His GraviTrap columns, see His GraviTrap/His GraviTrap Kit - Gravity-flow Purification.

IMAC Sepharose 6 Fast Flow, see IMAC Sepharose 6 Fast Flow Media/HiTrap IMAC FF Columns/HiPrep IMAC FF 16/10 Column/HiScreen IMAC FF Column, is the product of choice when charging the medium with different metal ions for optimization of purification protocols.

* For licensing information, see Legal Info.

TECHNICAL SPECIFICATIONS
Matrix	Highly crossed-linked 6% agarose, precharged with Ni2+
Average particle size	90 μm
Dynamic binding capacity*	Approx. 40 mg histidine-tagged protein/ml medium
Metal ion capacity	Approx. 15 μmol Ni2+/ml medium
Compatibility  during use	Stable in all commonly used buffers, reducing agents, denaturants, and detergents, see Introduction to Tagged Protein Purification for more information.
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 binding capacity is protein-to protein dependent † Ni2+-stripped medium

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