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This volume explores the latest methods used to study various aspects of TET proteins and their biology. Chapters in this book are divided into five parts. Part One describes technologies aimed at detecting and quantifying DNA methylation turnover using massively parallel sequencing, ELISA, and mass spectrometry approaches. Part Two looks at data analyses protocols for distinguishing acting versus passive DNA demethylation and estimation of 5mC and 5hmC levels. Part Three deals with a new topic that takes advantage of modified CRISPR/Cas9 genome editing systems to target DNA demethylation activity to genomic loci of interest. Part Four discusses protocols that detail how to purify TET proteins and unravel their protein interactions, and Part Five looks at the assessment of TET protein function and activity in vivo and in vitro. 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. Cutting-edge and thorough, TET Proteins and DNA Demethylation: Methods and Protocols is a valuable resource that aims to help research scientists at all levels working in the fields of DNA demethylation dynamics.Chapters 3, 7 and 17 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
This volume focuses on protein analysis, including a wide range of the use of mass spectrometry and other protein methods within neurobiological disciplines. Chapters cover topics such as cerebrospinal fluid (CSF) processing and biobanking; label-free quantitative proteomics; SWATH; top-down proteomics; and experimental strategies based on other ¿omics applied to CSF metabolome, lipidome, and microRNAome. 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.Cutting-edge and thorough, Cerebrospinal Fluid (CSF) Proteomics: Methods and Protocols is a valuable resource for graduate students and post-doctoral fellows interested in learning more about CSF proteotyping. It is also useful to established researchers seeking furtherinsight into this growing field.
This second edition volume expands on the previous edition with new sections describing the characterization of peptides bound to major histocompatibility complexes (MHC) on the surface of the cell. Chapters also cover topics such as using SERPA for antigen identification; antigen content of electroimmunoprecipitates; whole genome-phage display libraries; antigens in immune complexes; and immunoproteomic biomarkers. 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. Cutting-edge and thorough, Immunoprotemics: Methods and Protocols, Second Edition is a valuable resource that presents novice and expert researchers with techniques that are easily transferrable to laboratories and provides enhancedhands-on insights into this evolving field.
This volume explores the latest synthetic procedures for producing receptor-specific retinoids and rexinoids, molecular biology methods, and new technologies to demonstrate the therapeutic activities of molecules. The chapters in this book cover topics such as lentiviral delivery of shRNA constructs into acute promyelocytic leukemia cells for ATRA induced differentiation and autophagy; methods to analyze RAR signaling in colorectal cancer cells; differentiation of primary myoblasts by using RXR agonist; methodology for analyzing effects of retinoid treatment on nervous system development and larval swimming behavior; and protocols for assessment of autophagic flux in ATRA treated 2D and 3D breast cancer cultures. 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 troubleshootingand avoiding known pitfalls.Cutting-edge and comprehensive, Retinoid and Rexinoid Signaling: Methods and Protocols is a valuable resources for graduate students, postdoctoral fellows, and principal investigators who are interested in further exploring the signaling mechanisms of these molecules in their specific preclinical models.
This book brings together recent methods and theoretical approaches developed to dissect the activity and function of bacterial and eukaryotic Structural Maintenance of Chromosome (SMC) proteins. The protocols explore numerous subjects such as depletion systems to assess SMC function, genetic manipulation of SMC proteins, chromosomal, biochemical, and microscopy-based assays of SMC activity, as well as theoretical modeling of SMC activity. Written for the highly successful Methods in Molecular Biology series, 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 practical, SMC Complexes: Methods and Protocols will greatly aid researchers looking into the mechanism of action of SMC complexes on DNA.Chapters 10 and 19 areavailable Open Access under a CC BY 4.0 license via link.springer.com.
This detailed collection gathers both established and recent technical procedures to study the Endosomal Sorting Complex Required for Transport (ESCRT) complexes in a wide range of biological systems: Archaea, A. thaliana, U. maydis, S. cerevisiae, S. pombe, C. elegans, D. melanogaster, and mammalian cells. Opening with a section on imaging techniques, the book continues with chapters covering biochemical approaches presenting strategies for production and characterization of recombinant ESCRT proteins, or of specific ESCRT protein domains, as well as genetic and proteomic experimental approaches. Written for the highly successful Methods in Molecular Biology series, chapters include introduction 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 practical,The ESCRT Complexes: Methods and Protocols serves as a compact guide for researchers interested in establishing an integrated approach to investigate the ESCRT machinery functions in cell biology.
EPR spectroscopy is a versatile, nondestructive technique widely used in chemistry, biology, and physics. It detects molecules and materials with unpaired electrons making it a very selective technique that produces a wealth of information on such systems. Its high sensitivity makes it suitable in analyzing very small samples, single crystals, or reaction intermediates like radicals. This textbook takes a practical approach that introduces the basic concepts of EPR to sufficient detail to allow the reader to gain a basic knowledge of EPR and understand how experiments are carried out and how spectra are analyzed and interpreted. Many illustrative examples are included drawn from solid-state physics and bioinorganic chemistry. It is suitable as a short introduction for advanced undergraduate and beginning graduate students taking their first steps into EPR research.
Dieses Buch liefert einen sehr persönlichen Einblick in das Leben und Wirken von Felix Hoppe-Seyler. Er war durch seine Entdeckungen und seinen Einfluss auf die Entstehung eines neuen Forschungsgebietes, der Physiologischen Chemie oder Biochemie, eine der einflussreichsten Forscherpersönlichkeiten seiner Zeit. Hoppe-Seylers eigene Entdeckungen sind zahlreich. Sie schließen Erkenntnisse aus der Physiologie, der Pathologie und der Chemie ein. Im Mittelpunkt seiner Leistungen stehe aber die nie erlahmenden Bemühungen der Physiologischen Chemie, also der Wissenschaft von den chemischen Grundlagen der Biologie ihren Platz an den deutschen Universitäten zu erkämpfen. Seine Waffen waren die von ihm gegründete Zeitschrift für Physiologische Chemie und das Handbuch der Physiologischen- und Pathologischen Analyse. Georg Hoppe Seyler, ein Nachfahre, hat sich bemüht ein möglichst objektives, von überkommenen durch die Jahre veränderten Vorstellungen freies Bildauf der Grundlage von Briefen und Dokumenten zu zeichnen. Briefe des Nachlasses, Zitate von mit Felix Hoppe (-Seyler) arbeitenden Zeitgenossen die ihn als der Persönlichkeit und Wissenschaftler kennen lernten und seine Publikationen bilden den Hintergrund der kurzen Zusammenstellung.
With the interest in biomarkers available for diagnostic, therapeutic, prognostic and research applications growing rapidly, it is now expected that every pathologist should be aware of the correct application of immunohistochemistry in daily practice. This fully updated fourth edition provides practical guidance about preparation, key uses and pitfalls of antibodies for immunohistology in a unique A-Z format. Seventeen full colour infographics present detail about specific biomarkers, including FOXL2, GATA3 and SOX10, pulling out information about expression in tissues and prognosis. Expanded with new biomarkers and the removal of biomarkers that are no longer in common use, each entry in this concise new edition follows a standard structure for quick and easy access by busy readers.
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