推荐产品
包装
pack of 5 × 5 mL
制造商/商品名称
Cytiva 17-5286-01
参数
<20 mL/min flow rate
42 psi (H2O at room temperature.)
柱床尺寸
16 mm × 25 mm
柱床体积
5 mL
色谱柱内径
16 mm
基质
6% cross-linked agarose
粒径
45-165 μm
平均直径
90 μm
cleaning
2-14(Ni2+-stripped medium.)
工作范围
3-12(Ni2+-stripped medium.)
容量
~40 mg binding capacity (histidine-tagged protein/ml medium)(Protein binding capacity is protein-to-protein dependent.)
~40 mg binding capacity(histidine-tagged protein)
适用性
suitable for bioprocess medium
一般描述
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.
特点和优势
- 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
储存及稳定性
分析说明
法律信息
警示用语：
Warning
危险声明
储存分类代码
3 - Flammable liquids
法规信息
商品
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.
Follow established procedures for recombinant protein expression, growth, induction, and cell lysis optimization.
实验方案
HisTrap™ FF Kit purifies histidine-tagged proteins directly from cell lysates, minimizing sample pretreatment for rapid purification.
This page shows the purification strategy of capture, intermediate purification, and polishing (CIPP) in affinity chromatography using Cytiva products.
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.
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