产品名称
HisTrap™ Fast Flow Crude, Cytiva 17-5286-01, pack of 5 × 5 mL
form
prepacked column
crosslinking
cross-linked
packaging
pack of 5 × 5 mL
manufacturer/tradename
Cytiva 17-5286-01
storage condition
(20% Ethanol)
parameter
20 mL/min max. flow rate
5 bar max. pressure
5 mL/min flow rate
technique(s)
liquid chromatography (LC): suitable
bed volume
5 mL
column I.D. × L
16 mm × 25 mm
matrix
6% cross-linked agarose
matrix active group
Ni2+ ions
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)
separation technique
affinity
shipped in
ambient
storage temp.
2-30°C
Analysis Note
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
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.
Preparation Note
Legal Information
signalword
Warning
hcodes
存储类别
3 - Flammable liquids
法规信息
商品
Follow established procedures for recombinant protein expression, growth, induction, and cell lysis optimization.
This page describes manual and automated purification of histidine-tagged recombinant proteins using Cytiva products
This page shows the characteristics of Ni Sepharose, Ni Sepharose excel, TALON Superflow, and uncharged IMAC Sepharose products from Cytiva.
How to optimize purification of histidine-tagged proteins using Cytiva products.
实验方案
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 histidine-tagged proteins secreted into eukaryotic cell culture supernatants using HisTrap Excel from Cytiva.
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