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The ALFA-tag is a h...
The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications
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- Götzke, Hansjörg (author)
- NanoTag Biotechnologies GmbH
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- Kilisch, Markus (author)
- University Medical Center Göttingen
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- Martínez-Carranza, Markel (author)
- Stockholm University,Stockholms universitet,Institutionen för biokemi och biofysik
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- Sograte-Idrissi, Shama (author)
- University Medical Center Göttingen
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- Rajavel, Abirami (author)
- NanoTag Biotechnologies GmbH
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- Schlichthaerle, Thomas (author)
- Max Planck Institute of Biochemistry
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- Engels, Niklas (author)
- University Medical Center Göttingen
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- Jungmann, Ralf (author)
- Ludwig-Maximilian University of Munich,Max Planck Institute of Biochemistry
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- Stenmark, Pål (author)
- Stockholm University,Lunds universitet,Stockholms universitet,Institutionen för biokemi och biofysik,Lund University, Sweden,Strukturell biokemi,Forskargrupper vid Lunds universitet,Structural Biochemistry,Lund University Research Groups
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- Opazo, Felipe (author)
- NanoTag Biotechnologies GmbH,University Medical Center Göttingen
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- Frey, Steffen (author)
- NanoTag Biotechnologies GmbH
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(creator_code:org_t)
- 2019-09-27
- 2019
- English.
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In: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 10
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Abstract
Subject headings
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- Specialized epitope tags are widely used for detecting, manipulating or purifying proteins, but often their versatility is limited. Here, we introduce the ALFA-tag, a rationally designed epitope tag that serves a remarkably broad spectrum of applications in life sciences while outperforming established tags like the HA-, FLAG (R)- or myc-tag. The ALFA-tag forms a small and stable a-helix that is functional irrespective of its position on the target protein in prokaryotic and eukaryotic hosts. We characterize a nanobody (NbALFA) binding ALFA-tagged proteins from native or fixed specimen with low picomolar affinity. It is ideally suited for super-resolution microscopy, immunoprecipitations and Western blotting, and also allows in vivo detection of proteins. We show the crystal structure of the complex that enabled us to design a nanobody mutant (NbALFA(PE)) that permits efficient one-step purifications of native ALFA-tagged proteins, complexes and even entire living cells using peptide elution under physiological conditions.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Keyword
- Biochemistry
- biokemi
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Götzke, Hansjörg
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Kilisch, Markus
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Martínez-Carranz ...
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Sograte-Idrissi, ...
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Rajavel, Abirami
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Schlichthaerle, ...
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show more...
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Engels, Niklas
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Jungmann, Ralf
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Stenmark, Pål
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Opazo, Felipe
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Frey, Steffen
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show less...
- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Medical Biotechn ...
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and Medical Biotechn ...
- Articles in the publication
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Nature Communica ...
- By the university
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Stockholm University
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Lund University