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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

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About This Item

UNSPSC Code:
23151816
NACRES:
NA.56

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 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.cytiva.com.

Legal Information

FPLC is a trademark of Cytiva
HisTrap is a trademark of Cytiva
Sepharose is a trademark of Cytiva

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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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.

Follow established procedures for recombinant protein expression, growth, induction, and cell lysis optimization.

Protocols

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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