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Addition of imidazole during binding improves purity of histidine-tagged proteins

The purity of histidine-tagged proteins purified by metal chelate affinity chromatography can often be improved by optimizing the imidazole concentration in the sample and binding buffer to achieve a ...

An automated three-step purification of the VEGFR2 kinase domain using ÄKTAexplorer (from Life Science News, issue 6)

A histidine-tagged VEGFR2 kinase domain was expressed in baculovirus infected Trichoplusia ni cells and purified using a three-column step procedure (metal chelate affinity, anion exchange, gel ...

Automated tag removal using ÄKTAxpress (Life Science News 18, p.6-7)

ÄKTAxpress™ chromatography system, an integrated solution for high-throughput purification of His- and GST-tagged proteins, features methods for automated on-column removal of affinity tags. This ...

Effect of flow rate, molecular weight of (His)6-tagged proteins, and expression system on the performance of HisTrap HP columns (Life Sciences News 18, p 3-5)

(His) 6-tagged proteins were used in studies to evaluate the purification performance of HisTrap™ HP columns. (His) 6-tagged maltose-binding protein (MBP-[His]6,Mr ~43 000) expressed in E. coli was ...

Effective purification of histidine-tagged proteins using various metal ions (from Discovery Matters Issue 3, March 2006)

IMAC Sepharose™ High Performance is a new purification medium that can be charged with a metal ion of choice. Using this medium, we tested Cu2+, Zn2+, Co2+, and Ni2+ for the purification of a ...

Extended scope and quality of His-tagged recombinant protein purifications (from Life Science News Issue 16, February 2004)

The purification of His-tagged recombinant proteins by immobilized metal affinity chromatography (IMAC) is a popular and highly effective technique. Target products are recovered from cellular or ...

Gravity-flow purification of histidine-tagged proteins using Ni Sepharose 6 Fast Flow packed in PD-10 columns (from Discovery Matters Issue 1, July 2005)

A protocol for gravity-flow purification of a histidine-tagged green fluorescent protein (GFP-[His6]) using Ni Sepharose™ 6 Fast Flow packed in Empty Disposable PD-10 Columns is described. Using this ...

High-throughput purification of His- and GST-tagged proteins (from Life Science News Issue 17, August 2004)

ÄKTAxpress™ chromatography system is an integrated solution for high-throughput purification of His- and GST-tagged proteins. The system combines hardware, software and specially designed media to ...

Low-cost, automated serial purification of histidine-tagged proteins (Discovery Matters 6, Aug 2007)

Automated chromatographic systems are of great help to structural genomics programs involved in expressing and purifying numerous recombinant proteins. A low-cost and automated process, using ...

Monitoring expression of an E. coli membrane protein GFP-(His)8 fusion and purification using a combination of IMAC and gel filtration

An integral E. coli membrane protein GFP-(His)8-fusion was overexpressed, solubilized, and purified to homogeneity. The GFP-moiety allowed direct monitoring of the protein during all steps from ...

New gravity-flow column gives faster and simpler purification of histidine-tagged proteins

WebDocAbstract

Purification of a protein tagged with (His)6 at its N-terminus, C-terminus, and both N- and C-termini using different metal ions

Proteins can be labelled with histidine residues at either their N or C terminus. However, the position of the tag may affect protein behavior and, ultimately, its binding to metal ions. To ...

Scale-up and two-step purification of histidine-tagged proteins with Ni Sepharose 6 Fast Flow (from Discovery Matters Issue 1, July 2005)

Scaling up the purification of (histidine)6-tagged maltose binding protein (Mr 43 000) with prepacked HisTrap™ FF and HisPrep™ 16/10 FF columns (1-ml to 20-ml column formats) was achieved without ...

Screening buffer conditions using His MultiTrap FF 96-well filter plates for scale-up purification of a histidine-tagged protein on ÄKTAxpress (from Discovery Matters Issue 3, March 2006)

WebDocAbstract

Solutions for histidine-tagged protein purification (from Discovery Matters Issue 3, March 2006)

Immobilized Metal Affinity Chromatography (IMAC) is an excellent chromatography technique for purification of histidine-tagged proteins. Pure protein can be obtained from a cell extract in a single ...