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GE17-5268-01

Ni Sepharose® High Performance

GE Healthcare, 17-5268-01, pack of 25 mL

  •  NACRES NA.56

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Properties

Related Categories Affinity Chromatography, IMAC Matrices, Molecular Biology, Protein Chromatography, Proteomics More...
shelf life   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.
packaging   pack of 25 mL
mfr. no.   GE Healthcare, 17-5268-01
matrix   highly cross-linked 6% agarose
avg. part. size   34 μm
cleaning in place   2 - 14 (Ni2+-stripped medium.)
working range   3 - 12 (Ni²⁺-stripped medium.)
capacity   ≥40 mg binding capacity (histidine-tagged protein/ml medium)(Protein binding capacity is protein-to-protein dependent.)

Description

General description

Ni Sepharose® High Performance affinity media for high-resolution, high capacity purification of histidine-tagged proteins by immobilized metal ion affinity chromatography (IMAC).

Ni Sepharose® High Performance consists of highly cross-linked agarose beads to which a chelating group has been coupled. This chelating group is precharged with Ni2+, which will selectively retain proteins with exposed histidine groups.

The high stability and broad compatibility of Ni Sepharose® High Performance maintains biological activity and increases product yield, at the same time as it greatly expands the range of suitable operating conditions.

Features and Benefits

• High-performance purification of histidine-tagged proteins
• Negligible leakage of Ni2+ ions.
• High binding capacity, at least 40 mg/mL medium
• Compatible with a wide range of reducing agents, detergents, denaturants and other additives
• 34 μm bead size for improved resolution

Storage and Stability

Store at 4 to 30 °C (20% Ethanol)

Analysis Note

To view the Certificate of Analysis for this product, please visit www.gelifesciences.com.

Legal Information

IMAC Sepharose products, Ni Sepharose products and Fe Sepharose products
These products are sold under a license from Sigma-Aldrich under patent number EP 1276716 (Metal chelating compositions) and equivalent patents and patent applications in other countries.

Sepharose is a registered trademark of Cytiva

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Articles

Converting from Flow Velocity to Volumetric Flow Rates

Appendix 7, Extracted from Affinity Chromatography Vol. 1: Antibodies, GE Healthcare, 2016  
Keywords: Affinity chromatography, Chromatography

Protocols

Characteristics of Ni Sepharose, Ni Sepharose excel, TALON Superflow, and Uncharged IMAC Sepharose Products

Appendix 1, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016  
Keywords: Affinity chromatography, Buffers, Cell culture, Centrifugation, Chromatography, Detergents, Immobilization, Precipitation, Purification, Tagged proteins

Column Cleaning for Ion Exchange Chromatography and Chromatofocusing

Appendix 10, extracted from Ion Exchange Chromatography & Chromatofocusing, GE Healthcare, 2007
Keywords: Buffers, Chromatography, Detergents, Ion Exchange, Separation, Solvents

Column Packing and Preparation for Affinity Chromatography with Specific Groups of Biomolecules

Appendix 3, extracted from Affinity Chromatography Vol. 3: Specific Groups of Biomolecules, GE Healthcare, 2014
Keywords: Antimicrobials, Chromatography, Gene expression, Separation

Column Packing and Preparation

Appendix 6, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016  
Keywords: Affinity chromatography, Antimicrobials, Chromatography, Gene expression, Separation, Tagged proteins

Column packing and preparation for Ion Exchange Chromatography & Chromatafocusing

Appendix 3, extracted from Ion Exchange Chromatography & Chromatofocusing, GE Healthcare, 2007
Keywords: Antimicrobials, Chromatography, Gene expression, Ion Exchange, Purification, Separation

Desalting/Buffer Exchange and Concentration for Affinity Chromatography of Tagged Proteins

Desalting at laboratory scale is a well-proven, simple, and very fast method that will rapidly remove low molecular weight contaminants at the same time as transferring the sample into the desired bu...
Keywords: Absorption, Addition reactions, Affinity chromatography, Buffers, Centrifugation, Chromatography, Detergents, Dialysis, Fractionation, Ion Exchange, PAGE, Precipitation, Sample preparations, Separation, Size-exclusion chromatography, Tagged proteins

Handling Inclusion Bodies

Recombinant proteins are most often expressed in the intracellular space, but expression can also be controlled so that the protein is secreted into the periplasmic space or out into the culture medi...
Keywords: Affinity chromatography, Buffers, Chromatography, Detergents, Dialysis, Enzyme activity, Fractionation, Gene expression, Immobilization, Mass spectrometry, Nuclear magnetic resonance spectroscopy, PAGE, Purification, Reversed-phase chromatography, Sample preparations, Separation, Size-exclusion chromatography, Solvents, Sonication, Tagged proteins

Ion Exchange Chromatography Troubleshooting

If only certain peaks are of interest in this well-resolved separation, it may be advantageous to transfer to a step elution in order to save time and buffer. The rest of this section focuses on prac...
Keywords: Absorption, Adsorption, Buffers, Chromatography, Detergents, Digestions, Ion Exchange, PAGE, Precipitation, Sample preparations, Separation, Solvents

Minipreps using His SpinTrap and His SpinTrap Kit

His SpinTrap™ and His SpinTrap™ Kit are designed for efficient minipreps (small-scale purification) of histidine-tagged proteins directly from clarified or unclarified cell lysates. The columns may also ...
Keywords: Affinity chromatography, Buffers, Cell culture, Cell disruption, Centrifugation, Chromatography, Degradations, Gene expression, Homogenization, PAGE, Sample preparations, Sonication, Tagged proteins

Non-volatile and Volatile Buffer Systems

Appendix 2, extracted from Ion Exchange Chromatography & Chromatofocusing, GE Healthcare, 2007
Keywords: Chromatography, Ion Exchange

Optimizing Purification of Histidine-Tagged Proteins

Chapter 4, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016
Keywords: Affinity chromatography, Buffers, Cell disruption, Chromatography, Immobilization, Ion Exchange, PAGE, Purification, Separation, Sequencing, Size-exclusion chromatography, Tagged proteins

Performing High-Throughput Screening Using His MultiTrap HP and His MultiTrap FF 96-Well filter Plates

His MultiTrap™ HP and His MultiTrap™ FF are prepacked, disposable 96-well filter plates for reproducible, high-throughput screening of histidine-tagged recombinant protein expression. Typical applicat...
Keywords: Affinity chromatography, Biochemistry, Buffers, Cell disruption, Centrifugation, Chromatography, Detergents, Dot blot, Gene expression, PAGE, Precipitation, Size-exclusion chromatography, Tagged proteins

Performing a Cell Lysis of Recombinant Histidine-Tagged Proteins

For optimal conditions for growth, induction, and cell lysis of recombinant histidine-tagged proteins, please refer to established procedures. The following is a general procedure for cell lysis and ...
Keywords: Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Filtration, Homogenization, Precipitation, Sample preparations, Sonication, Tagged proteins

Practical Considerations for IEX Separation

This section covers detailed aspects of each step in an IEX separation, together with practical hints and tips to improve resolution and overall performance. In practice a separation can be summarize...
Keywords: Absorption, Buffers, Cell culture, Chromatography, Detergents, Filtration, Ion Exchange, Nucleic acid denaturation, PAGE, Phase transitions, Precipitation, Sample preparations, Separation, Solvents

Principles and Standard Conditions for Different Purification Techniques

Appendix 11, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016
Keywords: Affinity chromatography, Chromatography, Detergents, Ion Exchange, Ligands, Precipitation, Reversed-phase chromatography, Separation, Size-exclusion chromatography, Solvents, Tagged proteins

Purification of Protein A-Tagged Proteins

Chapter 8, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016
Keywords: Affinity chromatography, Chromatography, Fermentation, Gene expression, Growth factors, Ligands, Separation, Tagged proteins

Purification or Removal of Proteins and Peptides with Exposed Amino Acids: His, Cys, Trp, and/or with Affinity for Metal Ions

Extracted from Affinity Chromatography Vol. 3: Specific Groups of Biomolecules, GE Healthcare, 2014
Keywords: Adsorption, Buffers, Chromatography, Detergents, Immobilization, Ion Exchange, Ligands, Phase transitions, Precipitation, Reductions, Separation, Tagged proteins

Purification Using HisTrap HP and HisTrap FF

HisTrap HP and HisTrap FF are 1 ml and 5 ml HiTrap columns packed with Ni Sepharose High Performance or Ni Sepharose 6 Fast Flow, respectively. Sample application, washing,  and elution can be perfor...
Keywords: Affinity chromatography, Buffers, Chromatography, PAGE, Purification, Size-exclusion chromatography, Tagged proteins

Purification of Histidine-Tagged Recombinant Proteins Using Ni Sepharose High Performance

Ni Sepharose High Performance consists of highly cross-linked 6% agarose beads (34 µm) to which a chelating group has been immobilized and subsequently charged with Ni2+ ions. The chromatography medi...
Keywords: Affinity chromatography, Buffers, Cell disruption, Chromatography, Filtration, Homogenization, Immobilization, Purification, Sample preparations, Sonication, Tagged proteins

Sample Preparation for Affinity Chromatography in Specific Groups of Biomolecules

Appendix 1, extracted from Affinity Chromatography Vol. 3: Specific Groups of Biomolecules, GE Healthcare, 2014
Keywords: Absorption, Adsorption, Buffers, Cell culture, Centrifugation, Chromatography, Detergents, Dialysis, Electrophoresis, Filtration, Indicators, Nucleic acid denaturation, Precipitation, Sample preparations, Separation, Tagged proteins

Sample Preparation in Ion Exchange Chromatography

Appendix 1, extracted from Ion Exchange Chromatography & Chromatofocusing, GE Healthcare, 2007
Keywords: Absorption, Adsorption, Affinity chromatography, Amplification, Buffers, Cell culture, Centrifugation, Chromatography, Detergents, Dialysis, Electrophoresis, Filtration, Indicators, Ion Exchange, Nucleic acid denaturation, PAGE, Precipitation, Purification, Sample preparations, Separation

Selection of Purification Equipment

Appendix 4 extracted from Ion Exchange Chromatography & Chromatofocusing, GE Healthcare, 2007
Keywords: Chromatography, Ion Exchange, Purification, Separation

Sepharose High Performance: Purification with High Resolution

Use Sepharose High Performance media for purification of proteins, peptides or oligonucleotides.
Keywords: Buffers, Chromatography, Detergents, Filtration, High performance liquid chromatography, Ion Exchange, PAGE, Precipitation, Purification, Sample preparations, Separation, Solvents

Starting materials needed for the purification of integral membrane proteins for structural and functional studies

Extracted from Purifying Challenging Proteins - Principles and Methods, GE Healthcare, 2007
Keywords: Cloning, Detergents, Gene expression, Glycosylations, PAGE, Positron Emission Tomography, Separation, Tagged proteins

Tag Removal by Enzymatic Cleavage

In most cases, functional tests can be performed using the intact histidine-tagged protein. If removal of the tag is necessary, then procedures similar to GST tag removal can be followed, that is, sp...
Keywords: Affinity chromatography, Cell disruption, Centrifugation, Chromatography, Digestions, PAGE, Purification, Separation, Size-exclusion chromatography, Sonication, Tagged proteins

Troubleshooting Guide for Affinity Chromatography of Tagged Proteins

The troubleshooting guide below addresses problems common to the majority of purification products discussed in this chapter, as well as problems specific to a particular method. In the latter case, th...
Keywords: Addition reactions, Affinity chromatography, Buffers, Cell culture, Cell disruption, Centrifugation, Chromatography, Detergents, Fermentation, PAGE, Precipitation, Protein biosynthesis, Size-exclusion chromatography, Sonication, Tagged proteins, Western blot

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