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GE29-0486-31

HisTrap™ Fast Flow Crude

Cytiva 29-0486-31, pack of 1 mL

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About This Item

UNSPSC Code:
23151816
NACRES:
NA.56

packaging

pack of 1 mL

manufacturer/tradename

Cytiva 29-0486-31

availability

not available in North America

parameter

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)(Protein binding capacity is protein-to-protein dependent.)
 ~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).

Application

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.

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 ÄKTAdesign 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)

Other Notes

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

HisTrap is a trademark of Cytiva

Pictograms

Environment
GHS09

Signal Word

Warning

Hazard Statements

H226

Storage Class Code

3 - Flammable liquids

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

Performing a purification and on-column refolding of an insoluble histidine-tagged protein

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.

Performing a Cell Lysis of Recombinant Histidine-Tagged Proteins

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.

Protocols

Manual Purification Using HisTrap FF Crude Kit with a Syringe

This page shows how to purify histidine-tagged recombinant proteins manually with a syringe using HisTrap FF crude Kit from GE Healthcare.

Affinity Chromatography in a Purification Strategy (CIPP)

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.

Purification of Histidine-Tagged Proteins Secreted into Eukaryotic Cell Culture Supernatants Using HisTrap™ Excel

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

Purification from Unclarified Cell Lysate using HisTrap™ FF Crude

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

See All

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