HisTrap FF is a ready-to-use column, prepacked with precharged Ni Sepharose 6 Fast Flow for preparative purification of histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC).

High binding capacity, approx. 40 mg/ml medium.
Negligible leakage of Ni²⁺.
Prepacked columns offer reliable and convenient time-saving purification of histidinetagged recombinant proteins.
Compatible with a wide range of reducing agents, detergents, denaturants, 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 proteins. The negligible leakage of Ni²⁺ ions from the matrix ensures reliable capture of histidine-tagged proteins in repeated IMAC purifications

The high flow rates made possible by the Ni Sepharose 6 Fast Flow matrix make HisTrap FF columns well suited for small scale-up. Capacity can be increased by connecting columns in series. HiTrap IMAC FF is the product of choice when charging the medium with different metal ions for optimization of purification protocols.

HisTrap FF — Technical Specifications

Order Information	Use 20-40 mM imidazole in sample and binding buffers for highest purity.
Bed Volume	1 ml
Bed Dimensions	7 x 25 mm
Flow rate	<4 ml/min¹⁾
Storage Conditions	4 to 30°C, 20% Ethanol
Pressure Max. [Over the Packed Bed During Operation]	3 bar [0.3 MPa] (42 psi)
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.

Media

BioProcess Media	Yes
Average Particle Size	90 µm
Matrix	6% cross-linked agarose
Particle Size	45 µm-165 µm
Binding Capacity/ml Chromatography Medium	Approx. 40 mg histidine-tagged protein
Metal Ion Capacity	~15 µmol Ni₂₊/ml medium
pH stability Working Range	3-12¹⁾
pH stability Cleaning	2-14²⁾
Pressure/Flow Specification	Base matrix: 200-400 cm/h, 100 kPa, XK 50/60 column, bed height 25 cm
Storage Conditions	4 to 30°C, 20% Ethanol
Estimated Shelflife from Manufacture Date	3 years
Chemical Stability	Stable in commonly used aqueous buffers - 0.1 M HCl, 0.1 M NaOH, 8 M urea³⁾
¹⁾Ni²⁺-stripped medium.
²⁾Ni²⁺-stripped medium.
³⁾Ni²⁺-stripped medium.

Column

Complete Packsize	1 ml
Column i.d.	7 mm
Material [Column Hardware]	Polypropylene (PP)

Licensing

For licensing information, see the Licensing Statements page in the About Us section.

Animation
The principle of Affinity Chromatography Close

Brochure
Pure simplicity for tagged proteins Summary The use of recombinant proteins has increased greatly in recent years, as has the wealth of techniques and products used for their amplification and purification. The advantages of using a recombinant tagged protein to facilitate purification and detection are now widely recognized. In general, the first step in a protein expression workflow involves cloning of the target gene into an appropriate expression vector. This is followed by the transformation of the expression vector into a suitable host system for subsequent expression analysis. Close
ÄKTAxpress - Take advantage of your routines; be in two places at once Summary ÄKTAxpressTM is a complete, intelligent, and robust solution for multidimensional purifi cation of affinity-tagged proteins and antibodies. Automated multistep purifi cation provides the highest possible protein purity. ÄKTAxpress can run multiple samples providing both the required purity and high throughput. Robotic systems are limited to a single chromatography step and sacrifice purity to achieve high throughput. Now it is possible to purify proteins expressed at all levels, even proteins expressed at low levels can be purified. Dual-tagged proteins are purified easily and you have the option of on- or offcolumn tag cleavage. Close

Brochure

Pure simplicity for tagged proteins Summary

ÄKTAxpress - Take advantage of your routines; be in two places at once Summary

Data file
FAQ - Ni Sepharose Fast Flow, HisTrap Fast Flow, HisPrep 16/10 Fast Flow Close
Ni Sepharose 6 Fast Flow Summary Purifying histidine-tagged recombinant proteins by immobilized metal affinity chromatography (IMAC) continues to grow in popularity. Nickel (Ni2+) is the most used metal ion in such IMAC purifications. Ni Sepharose™ 6 Fast Flow is a BioProcess™ medium that combines the advantages of the Ni2+ with the recognized high flow rates of the Sepharose 6 Fast Flow matrix. The result is a IMAC medium that runs at high flow rates and that ensures smooth and trouble-free scale-up. As well as bulk media, Ni Sepharose 6 Fast Flow is supplied as HisPrep™ FF 16/10 and HisTrap™ FF prepacked columns for added speed, convenience, and reproducibility. Close

Data file

FAQ - Ni Sepharose Fast Flow, HisTrap Fast Flow, HisPrep 16/10 Fast Flow

Ni Sepharose 6 Fast Flow Summary

Handbook
Purifying Challenging Proteins - Principles and Methods Summary This handbook is intended for students and experienced researchers with an interest in the isolation of integral membrane proteins, multiprotein complexes, or in refolding proteins from inclusion bodies. The aim is to present tools, strategies, and solutions available to meet the purification challenges associated with these three classes of proteins. Close
Recombinant Protein Purification Handbook - Principles and Methods Summary This handbook is intended for those interested in the expression and purification of recombinant proteins. The use of recombinant proteins has increased greatly in recent years, as has the wealth of techniques and products used for their expression and purification. The advantages of using protein/peptide tag fused to the recombinant protein to facilitate its purification and detection is now widely recognized. In some cases, tags may improve the stability and solubility of recombinant proteins. The reader will be introduced to the initial considerations to be made when deciding upon host, vector, and use of a tagged or untagged protein. General guidelines for successful protein expression are also included. Advice is given on harvesting and extraction, handling of inclusion bodies, tag removal, and removal of unwanted salts and small molecules. Close
Strategies for protein purification Summary Strategies for Protein Purification is focused on chromatographic purification of proteins at research scale. Beginners can use the handbook to get a quick start when entering the field; experienced protein purifiers may also find valuable information. The handbook describes how to design a protein purification procedure, how to select and combine chromatography methods and purification formats, and how to monitor and evaluate protein purification. It gives an overview of the available methods and provides advice on how to avoid pitfalls in all operations, from initial sample preparation to final analysis of the purified protein. Close

Instruction/protocol
HisTrap FF, 1 ml and 5 ml Summary HisTrap™ FF is a ready to use column, prepacked with precharged Ni Sepharose™ 6 Fast Flow. This prepacked column is ideal for preparative purification of histidine tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC). HisTrap FF columns provide fast, simple, and easy separations in a convenient format, including a perfect start for scaling up. Ni Sepharose 6 Fast Flow has low nickel ion (Ni2+) leakage and is compatible with a wide range of additives used in protein purification. HisTrap FF columns can be operated with a syringe, peristaltic pump, or liquid chromatography system such as ÄKTAdesign™. Close

Instruction/protocol

HisTrap FF, 1 ml and 5 ml Summary

Magazine
Discovery Matters 1 Summary Includes: What’s new? Scale-up and two-step purification of histidine-tagged proteins with Ni Sepharose™ 6 Fast Flow Tandem affinity purification (TAP) of two subcomplexes of a transcription factor, TFIIH from Schizosaccharomyces pombe A comparative study of Superdex™ 75 10/300 GL versus TSK-GEL Super AW3000 for analytical gel filtration Automated identification and quantitation of proteins and peptides in mammalian samples using LC-MS/MS siRNA screening of the cell cycle with two dynamic GFP sensors SPA Imaging Charge-Based Binding Beads: A different approach to the capture of assay product High-throughput preclinical imaging Ettan™ DIGE: New protocol for selective labeling of cell surface proteins using CyDye™ DIGE Fluor minimal dyes Gravity-flow purification of histidine-tagged proteins using Ni Sepharose 6 Fast Flow packed in PD-10 columns Close
Discovery Matters issue 14 Summary Discovery Matters is a quarterly magazine from GE Healthcare with a wide range of articles covering topics relevant to the life science researcher. This issue (issue 14, October 2011) contains articles on: 1) Automated purification of antibody fragments using protein A and G prepacked gravity-flow columns 2) Multistep purification of tagged proteins for surface-plasmon resonance studies using ÄKTAxpress™ 3) The use of high-content analysis to develop a high-throughput wound healing assay for cancer research 4) Comparative performance of Amersham™ Hyperfilm™ Blue film for visualization of proteins The What’s New Section covers the major launches during Q2 and Q3 2011. Close

Magazine

Discovery Matters 1 Summary

Discovery Matters issue 14 Summary

Magazine article
Multistep purification of recombinant tagged proteins for SPR studies using ÄKTAxpress™ - article from Discovery Matters 14, October 2011 Summary Recombinant tagged DNaseE7 and NC10 scFv-Im7, used for the development of a novel protein capture assay based on surface plasmon resonance (SPR) technology, were purified on ÄKTAxpress™ system from E. coli lysates. Single-step methods were replaced by unattended multistep methods, incorporating multiple chromatography techniques: Affinity (IMAC or a customized affinity chromatography [AC] column), desalting (DS), ion exchange chromatography (IEX), and gel filtration (GF). An overnight dialysis step into a low pH buffer was successfully replaced by a DS step into the same buffer, automating the purification. These methods, incorporating operator predefined UV280 peak collection parameters, have enabled us to rapidly and reliably purify the recombinant tagged proteins. Close
Redesigned HiTrap fittings offer easier connection to ÄKTAdesign chromatography systems (Life Science News 18, p19) Summary The new inlet and outlet design of HiTrap columns eliminates the need for column connectors when used with ÄKTAdesign™ chromatography systems. Connection to a syringe, laboratory pump, or chromatography system such as FPLC™ System is also possible using the provided connectors. The guide on this page shows the different types of connector and explains their use. Close
Scale-up and two-step purification of histidine-tagged proteins with Ni Sepharose 6 Fast Flow (from Discovery Matters Issue 1, July 2005) Summary Scaling up the purification of (histidine)6-tagged maltose binding protein (Mr 43 000) with prepacked HisTrap™ FF and HisPrep™ 16/10 FF columns (1-ml to 20-ml column formats) was achieved without change in purity or loss of recovery. Moreover, a two-step purification of N-terminal (histidine)10-tagged Trx-P450 (Mr 133 200) using a HisTrap FF column for the first step and gel filtration for the second step is described. With this approach, the full-length target protein could be separated from two histidine-tagged truncated forms of the target protein. Close
Solutions for histidine-tagged protein purification (from Discovery Matters Issue 3, March 2006) Summary Immobilized Metal Affinity Chromatography (IMAC) is an excellent chromatography technique for purification of histidine-tagged proteins. Pure protein can be obtained from a cell extract in a single purification step. The (histidine)6 -tag has become the most used affinity tag due to its small size, strong metal ion binding, and ability to bind under denaturing conditions. IMAC with Ni2+ is the standard method for purifying histidine-tagged proteins, but in some cases other metal ions give the best results. GE Healthcare offers four media for IMAC in a variety of formats for efficient purification of histidine-tagged proteins. Ni Sepharose™ High Performance and Ni Sepharose 6 Fast Flow are precharged with Ni2+, while IMAC High Performance and IMAC Sepharose 6 Fast Flow are uncharged and can be used with a metal ion of your choice. Use this selection guide to choose the best product to meet your purification needs. Close

Material Safety Data Sheet
HisTrap FF, 5 ml, 5 x 5 ml Close

Poster
Automated multistep purification of histidine-tagged proteins from unclarified cell lysates Summary HisTrap™ FF crude is a ready to use column intended for purification of histidine-tagged proteins from unclarified cell lysates. No centrifugation or filtration of the samples is required before loading. The total purification time is thereby significantly reduced, which is an important advantage when working with many samples. In this study, ÄKTAxpress™, a chromatography system designed for high-throughput protein purification, was used for the purification of unclarified samples. HisTrap FF crude was used in the first purification step, followed by upto three additional chromatography steps. A comparison between unclarified sample purified on HisTrap FF crude and clarified sample purified on HisTrap FF is presented. Further, purification of a large volume of unclarified lysate as well as a large amount of target protein is shown. Close
Automated purification of dual-tagged proteins Summary Recombinant proteins can be designed to contain N- or C-terminal affinity tags. If including two different tags in a construct, the target protein can be purified utilizing different affinity media, producing a purer protein with a straightforward purification scheme. If different tags are positioned on each end of the protein, a dual-tagged purification scheme ensures that a full-length protein is the result. If cleavage sites are included in between the tag and the target protein, the affinity tag can be removed. In this study two comparisons are made. First, the same protein is purified using two different affinity columns in two different set-ups. For another dual-tagged protein, final purity is compared when using only one affinity column, relative to using two different affinity columns. Close
Use of IMAC magnetic beads for simplified detergent screening for optimization of purification workflows of histidine-tagged membrane proteins Summary Integral membrane proteins are main targets for development of new pharmaceuticals. Affinity purification is a powerful tool that has become the prime choice as capture step in membrane protein purification. Recombinant membrane proteins are commonly purified by use of polyhistidine tags and immobilized metal affinity chromatography (IMAC). The choice of detergent is the major key factor for successful solubilisation and further purifications of integral membrane proteins to avoid protein loss and inactivation. Therefore a detergent screen is often necessary to find the optimal detergent for each purpose. The use of the Membrane Protein Purification Kit allows for rapid, small-scale screening of optimal purification conditions for histidine-tagged membrane proteins. The kit contains IMAC magnetic beads pre-charged with Ni2+-ions (His Mag Sepharose™ Ni), selected detergents, buffers and an easy-to-follow protocol. Close

Selection guide
Affinity chromatography columns and media Summary This Selection Guide for affinity chromatography columns will help you find the right column or media. Areas: 1) group specific purification 2) isolation and purification of immunoglobulins 3) recombinant fusion proteins 4) coupling ligands 5) metal chelate chromatography Close
Guide sheet ÄKTA avant Summary A selection guide which shows prepacked columns designed to work with ÄKTA avant 25 Close
Guide sheet ÄKTApurifier Summary A selection guide which shows prepacked columns designed to work with ÄKTApurifier Close
Prepacked chromatography columns for ÄKTA™ design systems Summary Selection guide describing all the prepacked columns available from GE Healthcare for use with ÄKTA™ chromatography systems. Close
Prepacked columns designed to work with ÄKTAxpress Summary ÄKTAxpress™. Unattended purification of tagged proteins and antibodies. Prepacked columns designed to work with ÄKTAxpress. Close
Protein sample preparation Selection Guide Summary Almost all protein samples need further preparation after collection. The quality of sample preparation is crucial to get the best possible analytical results. This selection guide helps you to find the appropriate methods and products to obtain proteins for further studies in scales from nanograms to milligrams of purified protein. Three separate workflows are available for different sources of target protein; recombinant proteins, antibodies, and proteins from natural sources. These workflows are further refined and adapted to suit different number of samples, from a few to many such as in screening applications. Close
Total solutions for preparation of histidine-tagged proteins - Selection guide Summary Purification from unclarified lysates Batch/gravity flow purification Protein preps and mini-preps High-throughput screening Scale-up Close

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Accessories

Select to expand the desired collection of accessories.

Running HiTrap, HisTrap and GSTrap with a stand alone pump

Running HiTrap, HisTrap and GSTrap with a syringe

Running HiTrap, HisTrap and GSTrap with ÄKTAdesign systems

For accessories not displayed in this section, please contact your local GE Healthcare sales representative.

Running HiTrap, HisTrap and GSTrap with a stand alone pump

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#	Product Name	Product Code
1	Tubing Cutter	18-1112-46
1	Tubing, 2 m, i.d. 0.75 mm, o.d. 1/16"	18-1112-53
1	Tubing i.d. 0.5 mm, o.d. 1/16"	18-1113-68
1	Tubing, 2 m, i.d. 1.0 mm, o.d. 1/16"	18-1115-83
1	Tubing i.d. 0.25 mm, o.d. 1/16"	18-1120-95
2	HiTrap/HiPrep, 1/16", male connector for ÄKTAdesign	28-4010-81

Running HiTrap, HisTrap and GSTrap with a syringe

/gehcls_images/GELS/Images/TechSupportImages/Files/18114736_47.jpeg

#	Product Name	Product Code	Price	Qty
1	Connector 1/16" Male/Luer Female	18-1112-51

Running HiTrap, HisTrap and GSTrap with ÄKTAdesign systems

/gehcls_images/GELS/Images/TechSupportImages/Files/18114736_45.png

#	Product Name	Product Code
1	Tubing Cutter	18-1112-46
1	Tubing, 2 m, i.d. 0.75 mm, o.d. 1/16"	18-1112-53
1	Tubing i.d. 0.5 mm, o.d. 1/16"	18-1113-68
1	Tubing, 2 m, i.d. 1.0 mm, o.d. 1/16"	18-1115-83
1	Tubing i.d. 0.25 mm, o.d. 1/16"	18-1120-95
2	HiTrap/HiPrep, 1/16", male connector for ÄKTAdesign	28-4010-81
3	Fingertight Connector 1/16" Male for Tubing o.d. 1/16"	18-1112-55