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Brings together a number of core protocols concentrating on protein, carefully written and edited by experts, including: pulse-chase analysis to measure protein degradation; labeling a protein with fluorophores using NHS ester derivitization; immunoaffinity purification of proteins; and proteolytic affinity tag cleavage.

Linkers in Biomacromolecules, Volume 647 in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. New sections in this updated release include chapters devoted to ser-gly linkers + models, sergGly- EAAAK linkers, Effect of crowding on linker behavior, Protathether. Experimental approach to quickly test different linkers in fusion proteins, Linkers in enzyme fusions/cascade catalysis, Linker dependence of avidity in multivalent interactions between disordered proteins, ER/K ╬▒-helix linker, The effect of chirality and steric hindrance on intrinsic backbone conformational propensities: tools for protein design, Linker design in protein-based nanostructures, and much more. Linkers in Biomacromolecules, Volume 647 in the Methods in Enzymology series continues the legacy of this premier serial with quality chapters authored by leaders in the field. New sections in this updated release include chapters devoted to ser-gly linkers + models, sergGly- EAAAK linkers, Effect of crowding on linker behavior, Protathether. Experimental approach to quickly test different linkers in fusion proteins, Linkers in enzyme fusions/cascade catalysis, Linker dependence of avidity in multivalent interactions between disordered proteins, ER/K ╬▒-helix linker, The effect of chirality and steric hindrance on intrinsic backbone conformational propensities: tools for protein design, Linker design in protein-based nanostructures, and much more.Provides the authority and expertise of leading contributors from an international board of authorsPresents the latest release in the Methods in Enzymology series

Methods in Enzymology, Volume 646, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Chapters in this new release include Methods for Studying RNA condensation/granules in vitro, RNA Dynamics in Intracellular Condensates, Methods for Viscoelastic Characterization of Liquid and Gel Condensates, Incorporating Proteins into Complex Coacervates, Methods for Study of Liquid-Liquid Phase Coexistence in Proximity to Lipid Membranes, Preparation of and Solute Partitioning in Multiphase Coacervates, Reversible photocontrol of DNA coacervation, Enzymatic Control over Coacervation, and much more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology series

Methods in Enzymology, Volume 650 continues the legacy of this premier serial with quality chapters authored by leaders in the field. Chapters in this new release include Biophysical methods to study lanthanide-protein interactions, Genetically encoded sensors to study lanthanide biology, Spectrophotometric methods to determine the stability constants of lanthanide-macromolecule complexes, Lanthanide-based probes for amino acid modifications, In vitro selection and application of lanthanide-dependent DNAzymes, LRET biosensors for imaging protein interactions in living cells, Synthetic Modeling of the Structure and Function of the Lanthanide-Dependent, MDH Cofactor, EPR spectroscopy of lanthanides, Macromolecular crystallography for f-element complex characterization, and much more.Provides the authority and expertise of leading contributors from an international board of authorsPresents the latest release in the Methods in Enzymology series

mRNA 3'' End Processing and Stability, Volume 655 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of timely topics. Each chapter is written by an international board of authors.Provides the authority and expertise of leading contributors from an international board of authorsPresents the latest release in the Methods in Enzymology seriesUpdated release includes the latest information on mRNA 3'' End Processing and Stability

Ion Channels, Part C, Volume 653 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a variety of topics, including Nonsense suppression in ion channels, Engineering Ion Channels Using Protein Trans-splicing, Probing Ion Channel Neighborhoods Using APEX, STX based probes for NaVs, ANAP: a versatile, fluorescent probe of ion channel gating and regulation, High Throughput Screens for Small Molecule Ion Channel Modulators, Using toxins to study ion channels, Re/de-constructing ubiquitin regulation of ion channels, Tethered Peptide Toxins for Ion Channels, Voltage-Sensing Phosphatase Molecular Engineering, and more. Additional chapters cover Engineering excitable cells, Stretch and Poke Stimulation of Mechanically-Activated Ion Channels, Optical Control of STIM Channels, High Throughput Electrophysiological Evaluation of Mutant Ion Channels, Evaluating BEST1 Mutations in RPE Stem Cells, Long Read Transcript Profiling of Ion Channel Splice Variants, Permeation of Connexin Channels, Ratiometric pH indicator for melanosomes and lysosomes, and Ion channels in the epithelial cells of the choroid plexus.Provides the authority and expertise of leading contributors from an international board of authorsPresents the latest release in the Methods in Enzymology series

Ion Channels Part A, Volume 651 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a variety of new developments on the topic. Each chapter is written by an international board of authors.Provides the authority and expertise of leading contributors from an international board of authorsPresents the latest release in the Methods in Enzymology series

Ion Channels Part B, Volume 652 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of topics, including NMDAR, Pannexin, and CALHM, Making NaV1.4 and NaV1.7, TRPVs, Purification native nAChRs, GABAR Radu Aricescu, TRPV5/2, NaV1.5, KATP, TRPA1, TREK-1, SARS-CoV-2 3a ion channel, Ion channel conformational dynamics by encoded unnatural amino acid, Fluorescence lifetime measurement of absolute membrane potential, Fluorescent Toxins as Activity Sensors, FRET Analyses of Ion Channel Protein-Protein Interactions, Control of Ion Channel Gating with Photo-Switchable Tweezers, and Counting Subunits in Kv Channel Complexes.Provides the authority and expertise of leading contributors from an international board of authorsPresents the latest release in the Methods in Enzymology series

Antimicrobial Peptides, Volume 663 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on Unifying the classification of antimicrobial peptides in the Antimicrobial Peptide Database, Optimizing peptide library creation for PepSAVI-MS (RP libraries, etc.), Discovery of novel Antimicrobial peptides using BioProspecting, Screening for cysteine-stabilized scaffolds for developing protelytic-resistant AMPs, Exploring synergy and its role in antimicrobial peptide biology, Colorimetric assays for the rapid and high-throughput screening of antimicrobial peptide activity against diverse bacterial pathogens, and much more. Other chapters cover Liquid chromatography-mass spectrometry-based analysis of naturally occurring neuropeptide diastereomers, Multiplexed Quantitative Neuropeptidomics via DiLeu Isobaric Tagging, In vitro evaluation of antibiotic resistance via proteomics, Molecular networking-based strategies in mass spectrometry, Development of Macrocyclic antimicrobial peptides and peptoids, and a host of other timely topics. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in Methods in Enzymology serials Updated release includes the latest information on Antimicrobial Peptides

Pseudokinases, Volume 667, the latest release in the Methods in Enzymology serial, highlights new advances in the field with this new volume presenting interesting chapters, including the Production and Purification of the PEAK pseudokinases for structural and functional studies, Structural biology and biophysical characterization of Tribbles pseudokinases, Detecting endogenous TRIB protein expression and its downstream signaling, Analysis of human Tribbles 2 pseudokinase, Expression, purification and examination of ligand-binding to IRAK pseudokinases, Characterization of pseudokinase ILK-mediated actin assembly, Biochemical examination of Titin pseudokinase, Approaches to study pseudokinase conformations, CRISPR editing cell lines for reconstitution studies of pseudokinase function, and much more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in Methods in Enzymology serials Includes the latest information on Pseudokinases

B12 Enzyme, Volume 667 in the Methods in Enzymology serial, highlights new advances in the field with this new volume presenting timely, interesting chapters, including Part I: B12 Biosynthesis and B12 Chaperones, Probing the preference and regioselectivity of lower ligand activation and its role in cobamide diversity, Biosynthesis of corrinoids, The human B12 trafficking chaperones: CblC, CblD and ATR, A method for the efficient adenosylation of corrinoids, Production, purification and liposome reconstitution of cobamide synthase, Extraction and Purification of Cobamides from Bacterial Cultures, Part II: The many faces of B12-dependent enzyme reactions, Methylcobalamin-dependent methionine synthase, and much more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in Methods in Enzymology serials Includes the latest information on B12 Enzymes

Chemical Microbiology, Part B, Volume 665, the latest release in the Methods of Enzymology series, highlights new advances in the field, including comprehensive chapters on the Application of Antibiotic-derived Fluorescent Probes to Bacterial Studies, Metabolomic approaches for enzyme function and pathway discovery in bacteria, Adding a diazo-transfer reagent to culture to generate secondary metabolite probes for click chemistry, Customized Peptidoglycan Surfaces to Investigate Innate Immune Recognition via Surface Plasmon Resonance, Development and application of highly sensitive labeling reagents for amino acids, Bacterial Cell Wall Modification with a Glycolipid Substrate, and much more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology series Updated release includes the latest information on chemical microbiology