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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
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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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Benchtop Immobilized Metal Affinity Chromatography Reconstitution Assay Polyhistidine Tagged Metalloenzyme Undergraduate Laboratory Iron-binding Metalloenzymes Structure Function Teaching Laboratory Non-heme Iron-binding Enzymes Column Luer Lock Outlet Upper Cap Lower Bed Support Resin Particles Nickel-bound Nitriloacetic Acid Resin Ethanol Slurry Room Temperature Graduated Pipette Excess Storage Solution Pasteur Pipette Chilled Lysis Bind Buffer
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