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Key Documents

GE11-0004-58

HisTrap Fast Flow Crude

Cytiva 11-0004-58, pack of 5 × 1 mL

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About This Item

UNSPSC Code:
23151816
NACRES:
NA.56

packaging

pack of 5 × 1 mL

manufacturer/tradename

Cytiva 11-0004-58

parameter

flow rate
42 psi (H2O at room temperature.)

bed size

7 mm × 25 mm

bed volume

1 mL

column I.D.

7 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)
~40 mg binding capacity(histidine-tagged protein)

suitability

suitable for bioprocess medium

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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

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.
Store at 4 to 30 °C (20% Ethanol)

Analysis Note

To view the Certificate of Analysis for this product, please visit www.cytiva.com.

Other Notes

1)The pressure over the packed bed varies depending on a range of parameters such as the characteristics of the chromatography medium and the column tubing used.
The pressure over the packed bed varies depending on a range of parameters such as the characteristics of the chromatography medium and the column tubing used.

Legal Information

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

pictograms

EnvironmentFlame

signalword

WarningDanger

Hazard Classifications

Flam. Liq. 2

Storage Class

3 - Flammable liquids

wgk_germany

WGK 2


Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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Artículos

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 GE Healthcare.

This page shows a general procedure for cell lysis of recombinant histidine-tagged proteins using GE Healthcare products.

This page shows how to optimize purification of histidine-tagged proteins using GE Healthcare products.

Protocolos

AC separates proteins on the basis of a reversible interaction between a protein (or group of proteins) and a specific ligand coupled to a chromatography matrix. With such high selectivity and hence high resolution for the protein(s) of interest, purification levels in the order of several thousand-fold with high recovery of active material are achievable.

This page shows how to purify histidine-tagged proteins secreted into eukaryotic cell culture supernatants using HisTrap Excel from GE Healthcare.

This page shows how to purify histidine-tagged recombinant proteins from unclarified cell lysate using HisTrap FF crude from GE Healthcare.

This page shows how to purify protein A-tagged proteins using GE Healthcare products.

Ver todo

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