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Method Article
Purification of His-tagged membrane proteins from detergent-solubilized membranes
Dimitrios Fotiadis
Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, CH-3012 Bern, Switzerland
Corresponding Author
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This protocol describes a method to purify a His-tagged membrane protein form detergent-solubilized Escherichia coli (E. coli) membranes. Solubilized membranes are incubated with Ni-NTA that binds the His-tagged protein. After washing, His-tagged proteins are eluted with histidine. When starting with solubilized membranes or membrane pellets before detergent solubilization, protein purification takes half or one day, respectively.
Keywords
protein purification, membrane protein
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Associated Publications
Projection Structure of a Member of the Amino Acid/Polyamine/Organocation Transporter Superfamily
by F. Casagrande, M. Ratera, A. D. Schenk, +7
Journal Of Biological Chemistry (28 May, 2012)
Measuring substrate binding and affinity of purified membrane transport proteins using the scintillation proximity assay
by Daniel Harder and Dimitrios Fotiadis
Nature Protocols (03 August, 2012)
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This is a preprint. It has not completed peer review.
Method Article
Purification of His-tagged membrane proteins from detergent-solubilized membranes
Dimitrios Fotiadis
Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, CH-3012 Bern, Switzerland
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 07 Aug, 2012
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
This protocol describes a method to purify a His-tagged membrane protein form detergent-solubilized Escherichia coli (E. coli) membranes. Solubilized membranes are incubated with Ni-NTA that binds the His-tagged protein. After washing, His-tagged proteins are eluted with histidine. When starting with solubilized membranes or membrane pellets before detergent solubilization, protein purification takes half or one day, respectively.
Associated Publications
Projection Structure of a Member of the Amino Acid/Polyamine/Organocation Transporter Superfamily
by F. Casagrande, M. Ratera, A. D. Schenk, +7
Journal Of Biological Chemistry (28 May, 2012)
Measuring substrate binding and affinity of purified membrane transport proteins using the scintillation proximity assay
by Daniel Harder and Dimitrios Fotiadis
Nature Protocols (03 August, 2012)