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Sample and Buffer Preparation for SAXS

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Biological Small Angle Scattering: Techniques, Strategies and Tips

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1009))

Abstract

In this book chapter, a practical approach for conducting small angle X-ray scattering (SAXS) experiments is given. Our aim is to guide SAXS users through a three-step process of planning, preparing and performing a basic SAXS measurement. The minimal requirements necessary to prepare samples are described specifically for protein and other macromolecular samples in solution. We address the very important aspects in terms of sample characterization using additional techniques as well as the essential role of accurately subtracting background scattering contributions. At the end of the chapter some advice is given for trouble-shooting problems that may occur during the course of the SAXS measurements. Automated pipelines for data processing are described which are useful in allowing users to evaluate the quality of the data ‘on the spot’ and consequently react to events such as radiation damage, the presence of unwanted sample aggregates or miss-matched buffers.

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Acknowledgments

This work was supported by the Bundesministerium fur Bildung und Forschung (BMBF) project BIOSCAT, grant 05K12YE1 by iNEXT, grant number 653706, funded by the Horizon 2020 programme of the European Union and by a Human Frontier Science Program research grant, HFSP Ref: RGP0017/2012. M.A.G. was supported by an EMBL Interdisciplinary Postdoc Programme (EIPOD) and Marie Curie COFUND actions.

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Authors and Affiliations

  1. European Molecular Biology Laboratory (EMBL) Hamburg Outstation, DESY, Hamburg, Germany

    Melissa A. Graewert & Cy M. Jeffries

Authors
  1. Melissa A. Graewert
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  2. Cy M. Jeffries
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Corresponding author

Correspondence to Melissa A. Graewert .

Editor information

Editors and Affiliations

  1. G N Ramachandran Protein Center, CSIR Institute of Microbial Technology, Chandigarh, India

    Barnali Chaudhuri

  2. Structural Biology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain

    Inés G. Muñoz

  3. Center for Structural Molecular Biology and Neutron Scattering Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA

    Shuo Qian

  4. Center for Structural Molecular Biology and Neutron Scattering Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA

    Volker S. Urban

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© 2017 Springer Nature Singapore Pte Ltd.

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Graewert, M.A., Jeffries, C.M. (2017). Sample and Buffer Preparation for SAXS. In: Chaudhuri, B., Muñoz, I., Qian, S., Urban, V. (eds) Biological Small Angle Scattering: Techniques, Strategies and Tips. Advances in Experimental Medicine and Biology, vol 1009. Springer, Singapore. https://doi.org/10.1007/978-981-10-6038-0_2

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