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GE29-0486-31

HisTrap™ Fast Flow Crude

Cytiva 29-0486-31, pack of 1 mL

Synonym(s):

HIS chromatography column, HIS-tagged purification columns, IMAC column, Ni Sepharose 6 FF, immobilized metal ion chromatography column, prepacked column

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About This Item

NACRES:
NA.56
UNSPSC Code:
23151816

Key Documents

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Product Name

HisTrap™ Fast Flow Crude, Cytiva 29-0486-31, pack of 1 mL

form

prepacked column

crosslinking

cross-linked

packaging

pack of 1 mL

manufacturer/tradename

Cytiva 29-0486-31

storage condition

(20% ethanol)

parameter

1 mL/min flow rate
4 mL/min max. flow rate
5 bar max. pressure

technique(s)

liquid chromatography (LC): suitable

bed volume

1 mL

column I.D. × L

7 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

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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.

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

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).

Preparation Note

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)

Legal Information

HisTrap is a trademark of Cytiva

pictograms

Environment
GHS09

signalword

Warning

hcodes

H226

Storage Class

3 - Flammable liquids

Regulatory Information

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Articles

Cell Lysis of Histidine-Tagged Proteins

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

Manual and Automated Purification

This page describes manual and automated purification of histidine-tagged recombinant proteins using Cytiva products

Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose® Products

This page shows the characteristics of Ni Sepharose, Ni Sepharose excel, TALON Superflow, and uncharged IMAC Sepharose products from Cytiva.

Optimizing Purification of Histidine-Tagged Proteins

How to optimize purification of histidine-tagged proteins using Cytiva products.

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Protocols

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 Cytiva.

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 Cytiva.

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