title: Affinity purification of histidine-tagged proteins transiently produced in HeLa cells.
abstract: In order to produce eukaryotic proteins in a functional state, it is often necessary to use eukaryotic instead of prokaryotic expression systems. We have designed vectors which can be employed to express either N- or C-terminally histidine-tagged proteins in transiently transfected eukaryotic cells. The histidine tag allows the rapid enrichment of these proteins by metal chelate affinity chromatography in a native and functional state. Yields of up to 5 micrograms protein/5 x 10(7) cells were achieved.
authors.affiliation: Institute for Molecular Biology, Hannover Medical School, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Janknecht
authors.forename: A
authors.initials: A
authors.lastname: Nordheim
mesh_terms: D001483:Base Sequence; D002846:Chromatography, Affinity; D004268:DNA-Binding Proteins; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D005822:Genetic Vectors; D006367:HeLa Cells; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D050807:ets-Domain Protein Elk-1
chemical_list: D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D015682:Retroviridae Proteins, Oncogenic; D014157:Transcription Factors; D050807:ets-Domain Protein Elk-1; D006639:Histidine