ArticleBiological procedures online2016
Characterising protein/detergent complexes by triple-detection size-exclusion chromatography.
Article in Biological procedures online, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Fine-tuning the yeast GAL10 promoter and growth conditions for efficient recombinant membrane protein production and purification.Protein science : a publication of the Protein Society · 2025Article
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- Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis.International journal of molecular sciences · 2023Review
- Enabling online determination of the size-dependent RNA content of lipid nanoparticle-based RNA formulations.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences · 2021Article
- Article
- Preparation of a Deuterated Membrane Protein for Small-Angle Neutron Scattering.Methods in molecular biology (Clifton, N.J.) · 2021Article
- The Oligomeric State of the Plasma Membrane H⁺-ATPase fromMolecules (Basel, Switzerland) · 2019Article
- A Cholesterol Analog Induces an Oligomeric Reorganization of VDAC.Biophysical journal · 2019Article
- Conformational Dynamics Govern the Free-Energy Landscape of a Membrane-Interacting Protein.ACS omega · 2018Article
- Solution Structure of an Intramembrane Aspartyl Protease via Small Angle Neutron Scattering.Biophysical journal · 2018Article
- Saposin Lipid Nanoparticles: A Highly Versatile and Modular Tool for Membrane Protein Research.Structure (London, England : 1993) · 2018Article
- Discovery and characterization of stable and toxic Tau/phospholipid oligomeric complexes.Nature communications · 2017Article
- Quantification of Detergents Complexed with Membrane Proteins.Scientific reports · 2017Article
- Article
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3 authors.
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Abstract
backgroundIn vitro investigations of membrane proteins usually depend on detergents for protein solubilisation and stabilisation. The amount of detergent bound to a membrane protein is relevant to successful experiment design and data analysis but is often unknown. Triple-detection size-exclusion chromatography enables simultaneous separation of protein/detergent complexes and protein-free detergent micelles and determination of their molar masses in a straightforward and absolute manner. Size-exclusion chromatography is used to separate different species, while ultraviolet absorbance, static light scattering, and refractive index measurements allow molar mass determination of protein and detergent components.
resultsWe refined standard experimental and data-analysis procedures for challenging membrane-protein samples that elude routine approaches. The general procedures including preparatory steps, measurements, and data analysis for the characterisation of both routine and complex samples in difficult solvents such as concentrated denaturant solutions are demonstrated. The applicability of the protocol but also its limitations and possible solutions are discussed, and an extensive troubleshooting section is provided.
conclusionsWe established and validated a protocol for triple-detection size-exclusion chromatography that enables the inexperienced user to perform and analyse measurements of well-behaved protein/detergent complexes. More experienced users are provided with an example of a more sophisticated analysis procedure allowing mass determination under challenging separation conditions.
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