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GE17-5286-01

HisTrap Fast Flow Crude

GE Healthcare, 17-5286-01, pack of 5 × 5 mL

  •  NACRES NA.56

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Properties

Related Categories Additional Affinity Purification, Molecular Biology, Plant Biotechnology, Plant Proteomics, Protein Purification 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 5 × 5 mL
mfr. no.   GE Healthcare, 17-5286-01
parameter   <20 mL/min flow rate
  42 psi (H2O at room temperature.)
bed size   16 mm × 25 mm
bed volume   5 mL
column I.D.   16 mm
matrix   6% cross-linked agarose
particle size   45-165 μm
average diameter   90 μm
cleaning   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.)
  ~40 mg binding capacity(histidine-tagged protein)
suitability   suitable for bioprocess medium

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

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

FPLC is a trademark of Cytiva

HisTrap is a trademark of Cytiva

Sepharose is a registered trademark of Cytiva

Safety & Documentation

Safety Information

RIDADR 
UN1170 - class 3 - PG 3 - Ethanol, solution

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Protocols

Affinity Chromatography in a Purification Strategy (CIPP)

Chapter 9, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016
Keywords: Affinity chromatography, Chromatography, Ion Exchange, Ligands, Pharmaceutical, Reversed-phase chromatography, Size-exclusion chromatography, Tagged proteins

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

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

Manual and Automated Purification

Chapter 2, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016
Keywords: Affinity chromatography, Centrifugation, Chromatography, Gene expression, Tagged proteins

Manual purification using HisTrap FF crude Kit with a syringe

HisTrap™ FF crude Kit is designed for rapid and convenient manual purification of histidine-tagged proteins using premade buffers and a syringe. Histidine-tagged proteins can be purified directly from ...
Keywords: Affinity chromatography, Buffers, Cell disruption, Chromatography, Homogenization, PAGE, Precipitation, Sample preparations, Sonication, Tagged proteins, Western blot

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 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

Performing a purification and on-column refolding of an insoluble histidine-tagged protein from a 100 ml E. coli culture using HisTrap FF 1 ml with ÄKTAprime plus

This procedure uses a HisTrap FF 1 ml column but can also be used with a HisTrap HP 1 ml or a HisTrap FF crude 1 ml column. The procedure uses ÄKTAprime plus but can also be run on other ÄKTA systems...
Keywords: Buffers, Chromatography, Fractionation, Sonication, Tagged proteins

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 Membrane Proteins

Extracted from Purifying Challenging Proteins - Principles and Methods, GE Healthcare, 2007
Keywords: Adsorption, Buffers, Cell culture, Cell disruption, Characterizations, Chromatography, Degradations, Detergents, Filtration, Immobilization, Ion Exchange, Ligands, PAGE, Sample preparations, Separation, Sonication, 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 from unclarified cell lysate using HisTrap FF crude

HisTrap™ FF crude is a ready-to-use column, prepacked with Ni Sepharose® 6 Fast Flow, for purification of histidine-tagged recombinant proteins. After thorough cell disruption, it is possible to load ...
Keywords: Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Gene expression, Homogenization, PAGE, Precipitation, Size-exclusion chromatography, Sonication, Tagged proteins

Purification of Histidine-Tagged Proteins Secreted into Eukaryotic Cell Culture Supernatants Using HisTrap™ Excel

HisTrap excel 1 ml and 5 ml columns are ready-to-use IMAC columns prepacked with Ni Sepharose excel. The design of the columns in combination with the specific properties of the medium enables fast an...
Keywords: Affinity chromatography, Buffers, Cell culture, Chromatography, Degradations, Gene expression, PAGE, Tagged proteins

Purification of Histidine-Tagged Recombinant Proteins Using Ni Sepharose 6 Fast Flow

Ni Sepharose 6 Fast Flow consists of 90 µm beads of highly cross-linked agarose, to which a chelating ligand has been immobilized and subsequently charged with Ni2+ ions. The ligand density of Ni Sep...
Keywords: Affinity chromatography, Buffers, Cell disruption, Chromatography, Filtration, Homogenization, Immobilization, Ligands, Sample preparations, Sonication, 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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