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Journal / Biochemistry / Benchtop Immobilized Metal Affinity…
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Biochemistry

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JoVE Journal Biochemistry
Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory
 

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory

Article DOI: 10.3791/58012-v • 08:02 min • August 23rd, 2018
August 23rd, 2018 •
1, 1
1Department of Chemistry, Muhlenberg College

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Summary
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Here we present a protocol for the benchtop immobilized metal affinity chromatography purification and subsequent reconstitution of a polyhistidine tagged, non-heme iron binding dioxygenase suitable for the undergraduate teaching laboratory.

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Keywords: Benchtop Immobilized Metal Affinity Chromatography Polyhistidine Tagged Metalloenzyme Undergraduate Laboratory Non-heme Iron-binding Enzymes Nickel-bound Nitriloacetic Acid Resin Lysis Bind Buffer Cell-free Crude Extracts Flow-through Eluent
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