GE17-5286-01
HisTrap™ Fast Flow Crude
Cytiva 17-5286-01, pack of 5 × 5 mL
Synonym(s):
Histidine-tagged liquid chromatography column, FF crude columns, IMAC columns, protein purification columns
About This Item
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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
manufacturer/tradename
Cytiva 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
Related Categories
General description
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
Analysis Note
Legal Information
signalword
Warning
hcodes
Storage Class
3 - Flammable liquids
Certificates of Analysis (COA)
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Articles
This page shows how to perform a purification and on-column refolding of an insoluble his-tagged protein from an E. coli culture with HisTrap™ FF columns and ÄKTAprime from Cytiva.
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 sample preparation from bacterial cultures. Other established procedures may also work.
How to optimize purification of histidine-tagged proteins using Cytiva products.
This page describes principles and standard conditions for different purification techniques of histidine-tagged proteins using Cytiva products.
Protocols
This page shows how to purify histidine-tagged proteins secreted into eukaryotic cell culture supernatants using HisTrap Excel from Cytiva.
This page shows how to purify histidine-tagged recombinant proteins from unclarified cell lysate using HisTrap FF crude from Cytiva.
This page shows how to purify protein A-tagged proteins using Cytiva products.
This protocol shows how to remove histidine tags by enzymatic cleavage using Cytiva products.
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