Disorder for Dummies: Functional Mutagenesis of Transient Helical Segments in Disordered Proteins -- Computational Prediction of Intrinsic Disorder in Protein Sequences with the disCoP Meta-predictor -- Computational Prediction of Disordered Protein Motifs using SLiMSuite -- How to Annotate and Submit a Short Linear Motif to the Eukaryotic Linear Motif Resource -- Analyzing the Sequences of Intrinsically Disordered Regions with CIDER and localCIDER -- Exploring Protein Intrinsic Disorder with MobiDB -- An Easy Protocol for Evolutionary Analysis of Intrinsically Disordered Proteins -- Expression and Purification of an Intrinsically Disordered Protein -- Production of Intrinsically Disordered Proteins for Biophysical Studies; Tips and Tricks -- Recombinant Production of Monomeric Isotope-Enriched Aggregation-Prone Peptides: Polyglutamine Tracts and Beyond -- Cell-free Protein Synthesis of Small Intrinsically Disordered Proteins for NMR Spectroscopy -- Structural Analyses of Intrinsically Disordered Proteins by Small-Angle X-ray Scattering -- Determining Rg of IDPs from SAXS data -- Obtaining Hydrodynamic Radii of Intrinsically Disordered Protein Ensembles by Pulsed Field Gradient NMR Measurements -- Quantitative Protein Disorder Assessment using NMR Chemical Shifts -- Determination of pKa Values in Intrinsically Disordered Proteins -- Paris-DÉCOR: A Protocol for the Determination of Fast Protein Backbone Amide Hydrogen Exchange Rates -- Predicting Conformational Properties of Intrinsically Disordered Proteins from Sequence -- Enhanced Molecular Dynamics Simulations of Intrinsically Disordered Proteins -- Computational Protocol for Determining Conformational Ensembles of Intrinsically Disordered Proteins -- Computing, Analyzing and Comparing the Radius of Gyration and Hydrodynamic Radius in Conformational Ensembles of Intrinsically Disordered Proteins -- Binding Thermodynamics to Intrinsically Disordered Protein Domains -- Analysis of Multivalent IDP Interactions: Stoichiometry, Affinity, and Local Concentration Effect Measurements -- NMR Lineshape Analysis of Intrinsically Disordered Protein Interactions -- Measuring Effective Concentrations Enforced by Intrinsically Disordered Linkers -- Determining the Protective Activity of IDPs under Partial Dehydration and Freeze-thaw Conditions -- Screening Intrinsically Disordered Regions for Short Linear Binding Motifs -- Probing IDP Interactions with Membranes by Fluorescence Spectroscopy -- Protocol for Investigating the Interactions between Intrinsically Disordered Proteins and Membranes by Neutron Reflectometry -- Interactions of IDPs with Membranes Using Dark State Exchange NMR Spectroscopy -- Determination of Binding Kinetics of Intrinsically Disordered Proteins by Surface Plasmon Resonance -- Measuring and Analysing Binding Kinetics of Coupled Folding and Binding Reactions under Pseudo-first Order Conditions -- Understanding Binding Induced Folding by Temperature Jump -- Determining Binding Kinetics of Intrinsically Disordered Proteins by NMR spectroscopy -- Determination of Protein Phase Diagrams by Centrifugation -- In vitro Transition Temperature Measurement of Phase Separating Proteins by Microscopy -- Walking along a Protein Phase Diagram to Determine Coexistence Points by Static Light Scattering -- Expression and Purification of Intrinsically Disordered Aβ Peptide and Setup of Reproducible Aggregation Kinetics Experiment -- Measuring Interactions between Tau and Aggregation Inducers with Single Molecule Förster Resonance Energy Transfer -- Detection of Multisite Phosphorylation of Intrinsically Disordered Proteins using Phos-tag SDS-PAGE -- Multiple Site-specific Phosphorylation of IDPs Monitored by NMR -- Detection of Multisite Phosphorylation of Intrinsically Disordered Proteins using Quantitative Mass-Spectrometry -- Targeting an Intrinsically Disordered Protein by Covalent Modification -- Recording in-cell NMR-spectra in Living Mammalian Cells -- In-cell NMR of Intrinsically Disordered Proteins in Mammalian Cells -- Analyzing IDPs in Interactomes. .

The edition details methods to study intrinsically disordered proteins (IDPs) including recent topics such as extremely high-affinity disordered complexes, kinetics that evade established concepts, liquid-liquid phase separation, and novel disorder-driven allosteric mechanisms. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Intrinsically Disordered Proteins: Methods and Protocols aims to help scientists with different backgrounds to further their investigations into these fascinating and dynamic molecules. Chapter 24 is available open access under a CC BY 4.0 license via link.springer.com. Chapters "40 and 42 " are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.