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Superdex 75: Your Comprehensive Guide to Size-Exclusion Chromatography
Are you ready to unlock the power of precise protein separation and purification? Then understanding Superdex 75 is crucial. This comprehensive guide dives deep into the world of Superdex 75, a size-exclusion chromatography (SEC) resin widely used in biotechnology, pharmaceutical research, and various analytical applications. We'll cover its principles, applications, practical considerations, and troubleshooting tips, equipping you with the knowledge to master this powerful technique. Whether you're a seasoned researcher or just beginning your journey in protein analysis, this guide provides the essential information you need to effectively utilize Superdex 75.
Understanding Size-Exclusion Chromatography (SEC) and the Role of Superdex 75
Size-exclusion chromatography (SEC), also known as gel filtration chromatography, is a powerful liquid chromatography technique used to separate molecules based on their size and shape. The process involves passing a sample solution through a column packed with a porous stationary phase. Larger molecules, unable to penetrate the pores of the stationary phase, elute first, while smaller molecules, which can enter the pores, take longer to pass through the column and elute later.
Superdex 75 is a specifically designed SEC resin from Cytiva (formerly GE Healthcare) known for its excellent resolution, high capacity, and chemical stability. It's composed of cross-linked dextran beads, offering a wide range of pore sizes to effectively separate proteins within a specific molecular weight range. This makes Superdex 75 particularly suitable for purifying and analyzing proteins with molecular weights ranging from approximately 3 to 70 kDa.
Superdex 75: Applications Across Diverse Fields
The versatility of Superdex 75 extends across numerous scientific disciplines. Here are some key applications:
Protein Purification: Superdex 75 excels at separating proteins based on size, allowing researchers to isolate target proteins from complex mixtures such as cell lysates or fermentation broths. This is essential for downstream processing and characterization.
Molecular Weight Determination: By analyzing the elution profile of a protein on a Superdex 75 column, researchers can estimate its molecular weight. This information is critical for understanding protein structure and function.
Desalting and Buffer Exchange: Superdex 75 can efficiently remove salts, detergents, and other small molecules from protein samples, ensuring high sample purity before further analysis or application.
Oligonucleotide Purification: Superdex 75 can also be used to separate oligonucleotides based on size, making it useful in nucleic acid research.
Antibody Purification and Characterization: The accurate separation capabilities of Superdex 75 make it indispensable in antibody research, assisting in the purification and characterization of monoclonal and polyclonal antibodies.
Pharmaceutical Development: In drug discovery and development, Superdex 75 plays a crucial role in analyzing the purity and integrity of therapeutic proteins.
Practical Considerations for Using Superdex 75
Successfully employing Superdex 75 requires attention to several key factors:
Column Selection: Choosing the appropriate column size and dimensions is critical for optimal performance. Column length influences resolution, while diameter impacts sample loading capacity.
Mobile Phase Selection: The choice of buffer is crucial. It should be compatible with the sample, the resin, and the downstream applications. The ionic strength and pH of the buffer can significantly influence separation.
Sample Preparation: Proper sample preparation is essential. Ensure your sample is clarified and free from particulate matter to prevent column clogging. The sample volume should also be appropriate for the column size to achieve optimal resolution.
Flow Rate Optimization: The flow rate affects the resolution and runtime of the separation. An appropriate flow rate needs to be determined experimentally for optimal results. Too fast a flow rate may compromise resolution, while too slow a flow rate may increase runtime unnecessarily.
Column Equilibration and Regeneration: Proper equilibration of the column with the mobile phase before sample injection is critical for consistent results. Regeneration after use is essential to maintain column performance over multiple runs.
Data Analysis: Accurate data analysis requires careful calibration using molecular weight standards. This allows the accurate determination of molecular weights and aids in peak identification.
Troubleshooting Common Issues in Superdex 75 Chromatography
Despite careful planning, issues can arise during Superdex 75 chromatography. Here are some common problems and troubleshooting tips:
Poor Resolution: This may be caused by inappropriate flow rate, column overload, or improper equilibration. Try adjusting the flow rate, reducing sample volume, or re-equilibrating the column.
Column Clogging: This is often due to particulate matter in the sample. Ensure proper sample preparation including centrifugation or filtration.
High Back Pressure: This may indicate column clogging or problems with the pump system. Inspect the column for blockages and check the pump for proper function.
Unexpected Peak Elution: This may result from sample aggregation, degradation, or interaction with the column matrix. Investigate the sample's stability and consider alternative buffer conditions.
Low Recovery: Low recovery may stem from sample loss during sample preparation or nonspecific binding to the column matrix. Optimize sample preparation and consider alternative buffer conditions.
Ebook Outline: Mastering Superdex 75 Chromatography
Ebook Title: Mastering Superdex 75: A Practical Guide to Size-Exclusion Chromatography
Outline:
Introduction: What is Superdex 75? Why use it? Brief overview of the ebook content.
Chapter 1: Principles of Size-Exclusion Chromatography: Detailed explanation of SEC, including the separation mechanism, factors influencing separation, and terminology.
Chapter 2: Superdex 75 Resin Characteristics: Deep dive into Superdex 75's properties, including its composition, pore size distribution, and chemical stability.
Chapter 3: Applications of Superdex 75: Detailed exploration of the diverse applications of Superdex 75 across various scientific fields.
Chapter 4: Practical Guide to Superdex 75 Chromatography: Step-by-step instructions on column selection, sample preparation, mobile phase selection, flow rate optimization, and data analysis.
Chapter 5: Troubleshooting and Optimization: Comprehensive guide to troubleshooting common issues and optimizing the Superdex 75 chromatography process.
Conclusion: Recap of key concepts and future directions in SEC technology.
(Detailed expansion of each chapter would follow here, expanding on the points mentioned above in a far more detailed manner with additional figures, tables, and examples. This section would comprise the bulk of the ebook content, exceeding the current word count limitations.)
Frequently Asked Questions (FAQs)
1. What is the molecular weight range of Superdex 75? The optimal range is generally 3-70 kDa, but it can be used for slightly larger or smaller molecules depending on the specific application.
2. What type of buffer is suitable for Superdex 75? Many buffers can be used, but the choice should depend on the sample and downstream application. Phosphate buffers are frequently employed.
3. How do I determine the optimal flow rate for Superdex 75? The optimal flow rate needs to be determined experimentally, considering the column size and the resolution required.
4. How do I regenerate a Superdex 75 column? The regeneration procedure varies depending on the contaminants. Generally, it involves washing the column with cleaning solutions and re-equilibrating with the mobile phase.
5. What are the advantages of using Superdex 75 over other SEC resins? Superdex 75 offers excellent resolution, high capacity, and chemical stability.
6. Can Superdex 75 be used for desalting? Yes, Superdex 75 is effectively used for desalting and buffer exchange.
7. How do I interpret the elution profile obtained from Superdex 75 chromatography? The elution profile allows for the estimation of molecular weight and the assessment of sample purity. Calibration with molecular weight markers is essential.
8. What causes high back pressure during Superdex 75 chromatography? High back pressure may indicate column clogging, leaks in the system, or problems with the pump.
9. What are the limitations of Superdex 75 chromatography? Like all SEC techniques, Superdex 75 has limitations, primarily in resolving molecules with similar sizes and interacting molecules.
Related Articles
1. High-Performance Liquid Chromatography (HPLC): An overview of HPLC techniques and their application in protein analysis.
2. Protein Purification Techniques: A comparison of various protein purification methods, including affinity chromatography and ion-exchange chromatography.
3. Gel Filtration Chromatography Protocols: Detailed protocols for performing gel filtration chromatography using different resins.
4. Molecular Weight Determination Methods: An overview of various methods for determining protein molecular weight.
5. Antibody Engineering and Production: Discusses the importance of Superdex 75 in antibody purification and characterization.
6. Pharmaceutical Protein Analysis: An overview of the analytical techniques used in the pharmaceutical industry to characterize therapeutic proteins.
7. Chromatography Data Analysis: A guide to processing and interpreting chromatographic data.
8. Troubleshooting HPLC Systems: Comprehensive guide to diagnosing and resolving issues in HPLC systems.
9. Choosing the Right Chromatography Column: Factors to consider when selecting a chromatography column for a specific application.
| superdex 75: Column Handbook for Size Exclusion Chromatography Chi-san Wu, 1999-03-19 Column Handbook for Size Exclusion Chromatography is the first comprehensive reference to provide everything one needs to know about commercial analytical and preparative columns for size exclusion and gel filtration chromatography (SEC and GFC). SEC is now widely used as a quality assurance method in the polymer industry (both synthetic and biopolymers) to determine molecular weight and molecular weight distribution. The Handbook contains contributions from every column manufacturer around the world and from many experienced column users. It covers the technology, characterization, application, evaluation, maintenance, and quality control of analytical and preparative columns for SEC and GFC. Also included are columns for two closely related techniques, hydrodynamic chromatography and high osmotic pressure chromatography.Key Features* Evaluate and select columns with confidence for specific applications* Optimize separations and improve the ruggedness of analytical methods* Extend the service time of a column* Establish a quality-control program to ensure consistency in column performance* Avoid the expense of column damage or purchases that do not give the expected results |
| superdex 75: Protein Liquid Chromatography M. Kastner, 2000 Protein Liquid Chromatography is a handbook-style guide to liquid chromatography as a tool for isolating and purifying proteins, consisting of 25 individual chapters divided into three parts: Part A covers commonly-used, classic modes of chromatography such as ion-exchange, size-exclusion, and reversed-phase; Part B deals with various target protein classes such as membrane proteins, recombinant proteins, and glycoproteins; and Part C looks at various miscellaneous related topics, including coupling reaction, buffer solution additives, and software. The text as a whole can be viewed as a systematic survey of available methods and how best to use them, but also attempts to provide an exhaustive coverage of each facet. How to solve a specific problem using a chosen method is the overall essence of the volume. The principle philosophy of this compilation is that practical application is everything; therefore, both classical and modern methods are presented in detail, with examples involving conventional, medium- and high-pressure techniques. Over-exposure to history, concept, and theory has deliberately been avoided. The reader will find a wealth of tips and tricks from users for users, including advice on the advantages and disadvantages of each method. Easy-to-read sections on Getting started now and Where to go from here attempt to provide hands-on, fool-proof detailed practical procedures with complete and even standard model runs for any scientist or technician at work in this area. |
| superdex 75: The Protein Protocols Handbook John M. Walker, 2007-10-02 The Protein Protocols Handbook, Second Edition aims to provide a cross-section of analytical techniques commonly used for proteins and peptides, thus providing a benchtop manual and guide for those who are new to the protein chemistry laboratory and for those more established workers who wish to use a technique for the first time. All chapters are written in the same format as that used in the Methods in Molecular BiologyTM series. Each chapter opens with a description of the basic theory behind the method being described. The Materials section lists all the chemicals, reagents, buffers, and other materials necessary for carrying out the protocol. Since the principal goal of the book is to provide experimentalists with a full account of the practical steps necessary for carrying out each protocol successfully, the Methods section contains detailed st- by-step descriptions of every protocol that should result in the successful execution of each method. The Notes section complements the Methods material by indicating how best to deal with any problem or difficulty that may arise when using a given technique, and how to go about making the widest variety of modifications or alterations to the protocol. Since the first edition of this book was published in 1996 there have, of course, been significant developments in the field of protein chemistry. |
| superdex 75: Protein Phosphatases Susanne Klumpp, Josef Krieglstein, 2003 Protein phosphatases are a group of enzymes responsible for the dephosphorylation of various proteins and enzymes in a cell. This role is an extremely important one since protein phosphorylation and dephosphorylation is required for the regulation of a large number of cellular activities. Classification of Protein Phosphatases Analysis/Technology: Cell and Molecular Imaging Technology, Assay of Protein Phosphatases, MS and MNR, Genomics/Proteomics, cDNA Microarray Analysis Cellular Regulation: Substrates, Inhibitors, Regulation, Protein-Protein Interaction Biological Function: Antisense Studies, Transgenic and Knockout Animal Models in Vivo, Therapeutic Approaches |
| superdex 75: New and Emerging Proteomic Techniques Dobrin Nedelkov, Randall W. Nelson, 2008-02-04 Leading researchers and innovators describe in step-by-step detail the latest techniques that promise to significantly impact the practice of proteomics, as well as its success in developing novel clinical agents. The methods span the entire spectrum of top-down and bottom-up approaches, including microarrays, gels, chromatography, and affinity separations, and address every aspect of the human proteome, both quantitatively and qualitatively. The techniques of protein detection utilized are diverse and range from fluorescence and resonance light scattering to surface plasmon resonance and mass spectrometry. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. |
| superdex 75: Small Angle Scattering Part A: Methods for Structural Investigation , 2022-11-19 Small Angle Scattering, Part A: Methods for Structural Investigation, Volume 675 in the Methods in Enzymology series, highlights new advances in the field, with new chapters in this updated release including SAXS foundations and metrics, Contrast variation sample preparation protocols, experimental procedures, and rudimentary analysis, Molecular deuteration for neutron scattering, Planning, Executing and Assessing the Feasibility of SANS Contrast Variation Experiments, Technical considerations for small-angle neutron scattering from biological macromolecules, and Advanced sample environments and capabilities at our synchrotron X-ray beamline with example applications. Additional sections in the book cover SEC-SAXS-MALS data acquisition and processing pipeline at SIBYLS, SEC-SAXS: pros and cons, experimental set-up, examples and software developments, Radiation damage and sample economy for stopped-flow methods in the time regime of millisecond and above, Stopped-flow-time-resolved SAXS, Insights on Temp-jump, time-resolved SAXS, 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 - Includes the latest information on Small Angle Scattering: Methods for Structural Investigation |
| superdex 75: Protein Purification Jan-Christer Janson, 2011-03-22 The authoritative guide on protein purification—now completely updated and revised Since the Second Edition of Protein Purification was published in 1998, the sequencing of the human genome and other developments in bioscience have dramatically changed the landscape of protein research. This new edition addresses these developments, featuring a wealth of new topics and several chapters rewritten from scratch. Leading experts in the field cover all major biochemical separation methods for proteins in use today, providing professionals in biochemistry, organic chemistry, and analytical chemistry with quick access to the latest techniques. Entirely new or thoroughly revised content includes: High-resolution reversed-phase liquid chromatography Electrophoresis in gels Conventional isoelectric focusing in gel slabs and capillaries and immobilized pH gradients Affinity ligands from chemical and biological combinatorial libraries Membrane separations Refolding of inclusion body proteins from E. coli Purification of PEGylated proteins High throughput screening techniques in protein purification The history of protein chromatography |
| superdex 75: Branched-chain Amino Acids Robert Allison Harris, John Robert Sokatch, 2000 Volume 324 of Methods in Enzymology supplements Volume 166. It includes genetic information (cloning, gene expression) and information on human genetic diseases not available when Volume 166 was published. |
| superdex 75: A Text Book of PROTEIN PURIFICATION , |
| superdex 75: Growth Factors and Receptors Ian A. McKay, Kenneth D. Brown, 1998-07-02 Growth Factors and Receptors: A Practical Approach provides comprehensive protocols for studies of growth factors and their interactions with receptors. It covers a wide range from simple analytical techniques to sophisticated in vivo applications including: RT-PCR and immunocytochemistry for detection of growth factors and receptors; production and purification of recombinant growth factors and receptors; labelling of growth factors for binding studies; in vivo mutagenesis; the yeast two-hybrid assay of proteinprotein interactions; phage display of factors; application of factors to wound-healing processes using the gene gun; treatment of cancers with factor/toxin chimeras; and analysis of important factor domains using chimeric proteins. This book updates and extends the current literature and describes important novel approaches to the study of growth factors and their receptors, including the use of RNA aptamers as receptor antagonists, and the development of receptor superantagonists. It will be of tremendous value to both researchers and teachers, and, through an appendix that lists a large number of growth factors and receptors, will serve as a handy reference text. |
| superdex 75: Enzymes of Energy Technology Fraser Armstrong, 2018-11-30 Enzymes of Energy Technology, Volume 613 in the Methods in Enzymology series, highlights new advances in the field, with this updated volume presenting interesting chapters written by an international board of authors. Chapters include Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria, Production and manipulation of [NiFeSe]-hydrogenases for renewable hydrogen research, Hydrogen production by [FeFe]-hydrogenases, Production and properties of enzymes that activate and produce carbon monoxide, Recombinant [NiFe]-hydrogenases from E. coli, Working with nitrogenase, Oxygen--tolerant [NiFe]-hydrogenases, Cytoplasmic and Membrane Bound Hydrogenases from the hyperthermophile Pyrococcus furiosus, and more. Additional sections cover Enzymatic conversion of methane into useful chemicals, Production and investigations of trans-membrane electron transfer protein, Characterization of post-translational modifications in methyl-coenzyme M reductase in diverse methanogens by mass-spectrometry, Reductive activation of carbon dioxide by formate dehydrogenases, and Lytic polysaccharide monooxygenases in biofuel processing. 14. Production and manipulation of blue copper oxidases for technological applications Yasmina Mekmouche - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology series - Includes the latest information on the Enzymes of Energy Technology |
| superdex 75: Cyclic Nucleotide Signaling Xiaodong Cheng, 2015-05-07 Showcasing the recent progresses of the field, Cyclic Nucleotide Signaling covers the major tools and methodologies used in various areas of research. The majority of the chapters are protocol oriented, with the goal of providing clear directions for laboratory use. Students and investigators new to the field will find this book particularly inform |
| superdex 75: Microspheres and Microcapsules in Biotechnology Guanghui Ma, Zhi-Guo Su, 2013-01-29 Microspheres and microcapsules have very broad applications in various fields, especially in those of biotechnology and biopharmaceuticals, as targeting drug-delivery carriers, separation media for protein, peptide, DNA, and so forth. It is a big challenge to design and prepare microspheres and microcapsules of different sizes and structures from v |
| superdex 75: The Eukaryotic RNA Exosome John LaCava, Štěpánka Vaňáčová, 2019-11-25 This volume provides a cross-section of RNA exosome research protocols, applied to a diversity of model organisms. Chapters guide readers through methods that e.g. delineate eukaryotic exosomes’ origins in prokaryotes, probe its RNA substrates, adapter complexes and macromolecular interaction of networks, and establish critical structural-function relationships. 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, The Eukaryotic RNA Exosome: Methods and Protocols aims to ensure successful results in the further study of this vital field. |
| superdex 75: RNA Purification and Analysis Douglas T. Gjerde, Lee Hoang, David Hornby, 2009-07-10 This first book on the market covers the many new and important RNA species discovered over the past five years, explaining current methods for the enrichment, separation and purification of these novel RNAs. Building up from general principles of RNA biochemistry and biophysics, this book addresses the practical aspects relevant to the laboratory researcher throughout, while discussing the performance and potential problems of the methods discussed. An appendix contains a glossary with the important terms and techniques used in RNA analysis. By explaining the basic and working principles of the methods, the book allows biochemists and molecular biologists to gain much more expertise than by simply repeating a pre-formulated protocol, enabling them to select the procedure and materials best suited to the RNA analysis task at hand. As a result, they will be able to develop new protocols where needed and optimize and fine-tune the general purpose standard protocols that come with the purification equipment and instrumentation. |
| superdex 75: Gold and Silver Staining Gerhard W. Hacker, Jiang Gu, 2002-04-17 Immunogold silver staining is one of the most sensitive techniques available for visualizing the location of antibodies and nucleotide probes that have been bounded to specific antigens or to nucleotide sequences. As gold and silver staining continues to advance research in molecular morphology, this book presents the information you need to know a |
| superdex 75: Enzymology and Molecular Biology of Carbonyl Metabolism 10 H. Weiner, 2001-05-14 The largest collection of articles on the three major gene families, this work ranges from enzymology to molecular biology to physiological implications. The three gene families are related in that the enzymes catalyse the NAD(P) dependent oxidation or reduction of carbonyl containing substrates. The substrates are important in diverse areas such as alcoholism, diabetes and cancer related problems as well as simple detoxification. The scope of the chapters, contributed by leading international scientists, is wide and covers gene regulation to enzyme mechanisms and protein structure. This is the only publication dealing in such depth with just three gene families. An important reference for researchers in toxicology and molecular biology. |
| superdex 75: Purification and characterisation of O-demethylase from Acetobacterium dehalogenans Franz Kaufmann, 1998 |
| superdex 75: Hplc Of Biological Macro- Molecules, Revised And Expanded Karen M. Gooding, Fred E. Regnier, 2002-01-08 Completely revised to reflect the innovations in HPLC from the past decade, this authoritative reference presents practical strategies for the evaluation and analysis of proteins, peptides, and polynucleotides. Offering class-specific applications for the characterization and fractionation of biological macromolecules, the book contains material on organic supports, size exclusion, ion exchange, hydrophobic interaction, and metal interaction chromatography. Leading experts summarize specialized detection systems, provides discussions on the chemical and biological properties of specific biomolecules, include detailed guidelines for the development of analytical techniques, and more. |
| superdex 75: Placenta and Trophoblast Michael J. Soares, Joan S. Hunt, 2008-02-02 The aim of the two-volume set of Placenta and Trophoblast: Methods and Protocols is to offer contemporary approaches for studying the biology of the placenta. The chapters contained herein also address critical features of the female organ within which the embryo is housed, the uterus, and some aspects of the embryo–fetus itself, particularly those of common experimental animal models. In keeping with the organization used effectively in other volumes in this series, each chapter has a brief introduction followed by a list of required items, protocols, and notes designed to help the reader perform the expe- ments without difficulty. In both volumes, sources of supplies are given and illustrations highlight particular techniques as well as expected outcomes. A key aspect of these volumes is that the contributors are at the forefronts of their disciplines, thus ensuring the accuracy and usefulness of the chapters. Placenta research has progressed rapidly over the past several decades by taking advantage of the technical advances made in other fields. For example, the reader will note that many techniques, such as reverse transcriptase po- merase chain reaction, northern and western blotting, microarray analyses and in situ hybridization experiments, are routinely used for dissecting a wide range of experimental questions. Protein analysis and functional experiments on tissues and cells that comprise the maternal–fetal interface benefit from studies in en- crinology, immunology, and developmental biology. These volumes also present new ideas on investigating gene imprinting and gene transfer via viral vectors. |
| superdex 75: Oxygen/Nitrogen Radicals: Cell Injury and Disease Val Vallyathan, Vince Castranova, Xianglin Shi, 2012-12-06 A veritable mountain of literature has been published showing the causal relationship of reactive oxygen/nitrogen species in human disease conditions, and there has been an explosion in the understanding of oxidative stress, the protective role of antioxidants and molecular events involved in the regulation of transcription, editing, and translation of key events leading to disease processes. Strategies need to be developed for prevention of diseases by allowing scientists and clinicians to obtain information on new and emerging advances. The molecular mechanisms involved in several diseases including Alzheimer's disease, atherosclerosis, diabetes, arthritis, and Parkinson's disease, as well as disorders of the eye, skin, cardiac, and pulmonary systems are discussed in this volume, along with scientific evidence supporting the value of dietary supplementation with antioxidants in the prevention of cellular damage leading to chronic disease. Special in vivo techniques are also discussed at length, along with the role of molecular studies in human risk assessment. |
| superdex 75: Chromatography Leonardo Calderon, 2012-10-24 Nowadays, Chromatography is the most versatile and widespread technique employed in modern chemical analysis and plays a vital role in the advancement of chemistry, biology, medicine and related fields of research. Because of the inherent simplicity and ease of operation, it can be used together with a wide range of detection systems, including electrochemical, photometric and mass spectrometry, being an invaluable laboratory tool for the separation and identification of compounds. The purpose of this book is not only to present the latest state and development tendencies of chromatography, but to bring the reader useful information on separation sciences to enable him to use chromatography on his research field. Taking into account the large amount of knowledge about chromatography theory and practice presented in the book, it has three major parts: applications, theory and sample preparation. The book is also intended for both graduate and postgraduate students in fields such as chemistry, biology, biotechnology, forensic, medicine, pharmacology and engineering, and as a reference for professionals and practitioners. |
| superdex 75: Histone Methyltransferases Raphaël Margueron, Daniel Holoch, 2022-06-22 This volume provides methods used to investigate histone methyltransferase function. Chapters guide readers through a comprehensive set of approaches that detail phylogenetic diversity, histone demethylase activities in vitro, generating chromatin substrates, auto-methylation, quantification of metabolites, protein purification, crystallization, X-ray structure, cryogenic electron microscopy, assessing genome-wide patterns, CUT&Tag in mouse embryonic tissues, chemical biology approaches, peptide SPOT arrays, nascent chromatin capture, ectopic protein tethering, computational models, and development of methyltransferase inhibitors. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Histone Methyltransferases: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge. |
| superdex 75: Streptococci and the Host Thea Horaud, Anne Bouvet, Roland Leclercq, Henri de Montclos, Michel Sicard, 2013-11-08 Streptococci and enterococci are the etiologic agents of infectious diseases that rank among the most severe in human pathology. The diagnosis, antibiotherapy, and prevention of the streptococcal diseases have improved considerably. However, the reemergence of severe streptococcal and enterococcal diseases constitutes a growing public health con cern, which remains open to scientific and medical debate. The XIII'h Lancefield International Symposium on Streptococci and Streptococcal Diseases, held at Institut Pasteur, Paris, France, September 16---2el, 1996, attracted 505 par ticipants from 43 countries. Twenty-two percent of the participants were students, a clear sign of the intense interest in this field. Of the 390 presentations made at the symposium, 260 were submitted as manuscripts for the Proceedings; we have included 249 of these in this volume. This symposium provided a forum for the presentation of the most recent findings and approaches to understanding several important fields, such as new aspects of infec tion, bacteria~host interactions, epidemiology, and molecular genetics of streptococci and enterococci. Over the last three years, the study of these subjects has expanded as increas ingly sophisticated methods of molecular analysis have been applied to investigate the bi ology of pathogenic streptococci and enterococci. Virulence, vaccine strategies, genetics, antibiotic resistance, epidemiology, and immunology are now being examined through the lens of molecular biology. The application of recently developed techniques to this field will continue to yield insight into the mechanism by which these organisms cause disease. |
| superdex 75: Uronic Acids—Advances in Research and Application: 2012 Edition , 2012-12-26 Uronic Acids—Advances in Research and Application: 2012 Edition is a ScholarlyPaper™ that delivers timely, authoritative, and intensively focused information about Uronic Acids in a compact format. The editors have built Uronic Acids—Advances in Research and Application: 2012 Edition on the vast information databases of ScholarlyNews.™ You can expect the information about Uronic Acids in this eBook to be deeper than what you can access anywhere else, as well as consistently reliable, authoritative, informed, and relevant. The content of Uronic Acids—Advances in Research and Application: 2012 Edition has been produced by the world’s leading scientists, engineers, analysts, research institutions, and companies. All of the content is from peer-reviewed sources, and all of it is written, assembled, and edited by the editors at ScholarlyEditions™ and available exclusively from us. You now have a source you can cite with authority, confidence, and credibility. More information is available at http://www.ScholarlyEditions.com/. |
| superdex 75: Class 1 Oxidoreductases XII Antje Chang, Dietmar Schomburg, Ida Schomburg, 2008-01-23 The Springer Handbook of Enzymes provides concise data on some 5,000 enzymes sufficiently well characterized – and here is the second, updated edition. Their application in analytical, synthetic and biotechnology processes as well as in food industry, and for medicinal treatments is added. Data sheets are arranged in their EC-Number sequence. The new edition reflects considerable progress in enzymology: the total material has more than doubled, and the complete 2nd edition consists of 39 volumes plus Synonym Index. Starting in 2009, all newly classified enzymes are treated in Supplement Volumes. |
| superdex 75: Riboswitch Discovery, Structure and Function , 2014-11-21 This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in riboswitch discovery and validation, synthesis and sample prep methods for large RNAs, riboswitch structure and function methods, folding pathways and dynamics, and ligand interactions and thermodynamics. - Continues the legacy of this premier serial with quality chapters authored by leaders in the field - Covers research methods in riboswitch discovery, structure and function - Contains sections on such topics as riboswitch discovery and validation, synthesis and sample prep methods for large RNAs, riboswitch structure and function methods, folding pathways and dynamics, ligand interactions and thermodynamics |
| superdex 75: Biomass, Biofuels, Biochemicals Ram Sarup Singh, Reeta Rani Singhania, Ashok Pandey, Christian Larroche, 2019-01-31 Biomass, Biofuels and Biochemicals: Advances in Enzyme Technology provides state-of-the-art information on the fundamental aspects and current perspectives in enzyme technology to graduate students, postgraduates and researchers working in industry and academia. The book provides information about the use of enzyme technology as an important tool for biotechnological processes, including food, feed, fuels, textiles, paper, energy and environmental applications. The search for improvements in existing enzyme-catalyzed processes dictates the need to update information on various enzyme technologies. The book gives a snapshot of current practice and research in the area of enzyme technology. - Includes current and emerging technologies for the development of novel enzyme catalysis - Outlines immobilized enzymes and their implications - Refers to enzymes as diagnostic tools - Includes metabolic engineering principles for improving industrial enzymes |
| superdex 75: Handbook Of Size Exclusion Chromatography And Related Techniques Chi-San Wu, 2003-11-04 Documenting critical advances in this rapidly evolving field, the Second Edition highlights the need for new applications and technologies that assist in the determination of molecular weight and molecular weight distributions of polymers in an accurate, efficient manner. This volume presents the latest findings from a international team of specialists and continues to inspire and extend practical applications of size exclusion chromatography (SEC). It includes six new chapters covering high-speed size exclusion chromatography, SEC of low molecular weight materials, and the extended family of techniques, from two-dimensional liquid chromatography to high osmotic pressure chromatography. |
| superdex 75: Molecular Characterization of Autophagic Responses Part A , 2017-02-18 Molecular Characterization of Autophagic Responses, Part A, presents a collection of methods for the qualitative and quantitative evaluation of virtually all the morphological, biochemical, and functional manifestations of autophagy, in vitro, ex vivo and in vivo, in organisms as distant as yeast and man. Autophagy is an evolutionarily conserved mechanism for the lysosomal degradation of superfluous or dangerous cytoplasmic entities, and plays a critical role in the preservation of cellular and organismal homeostasis. Monitoring the biochemical processes that accompany autophagy is fundamental for understanding whether autophagic responses are efficient or dysfunctional. - Offers a detailed overview of the protocols used to study autophagy and various aspects of autophagic responses - Written in an accessible style by renowned experts in the field |
| superdex 75: Globins and Other Nitric Oxide-Reactive Proteins, Part A Robert K. Poole, 2011-09-02 The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 400 volumes (all of them still in print), the series contains much material still relevant today—truly an essential publication for researchers in all fields of life sciences. Methods in Enzymology is now available online at ScienceDirect — full-text online of volumes 1 onwards. For more information about the Elsevier Book Series on ScienceDirect Program, please visit: http://www.info.sciencedirect.com/bookseries/This volume features methods for the study of globin and other nitric oxide-reactive proteins. |
| superdex 75: Lipidomics and Bioactive Lipids: Lipids and Cell Signaling , 2007-11-12 This volume in the well-established Methods in Enzymology series features methods for the study of lipids using mass spectrometry techniques. Articles in this volume cover topics such as Phospholipase A1 assays using a radio-labeled substrate and mass spectrometry; Real-time Cell Assays of Phospholipases A2 Using Fluorogenic Phospholipids; Analysis and Pharmacological Targeting of Phospholipase C â interactions with G proteins; Biochemical Analysis of Phospholipase D.; Measurement of Autotaxin/Lysophospholipase D Activity; Platelet-Activating Factor; Quantitative measurement of PtdIns(3,4,5)P3; Measuring Phosphorylated Akt And Other Phosphoinositide 3-Kinase-Regulated Phosphoproteins In Primary Lymphocytes; Regulation of Phosphatidylinositol 4-Phosphate 5-Kinase activity by partner proteins; Biochemical Analysis of Inositol Phosphate Kinases; Analysis of the phosphoinositides and their aqueous metabolites; Combination of C17-sphingoid base homologues and mass spectrometry analysis as a new approach to study sphingolipid metabolism; Measurement of mammalian sphingosine-1-phosphate phosphohydrolase activity in vitro and in vivo; A rapid and sensitive method to measure secretion of sphingosine-1-phosphate; Ceramide Kinase and Ceramide-1-Phosphate; Measurement of Mammalian Diacylglycerol Kinase Activity in vitro and in Cells; Lipid Phosphate Phosphatases from Saccharomyces cerevisiae. |
| superdex 75: Casein kinases in human diseases Andrea Venerando, Giorgio Cozza, Victor Bustos, Lorenzo Alberto Pinna, 2023-01-06 |
| superdex 75: Selenium Victor R Preedy, 2015-08-10 Although toxic in large doses, selenium is an essential trace mineral in the animal diet and in some plants. It has a role in making antioxidant enzymes and a particular role in the functioning of the thyroid gland. This volume examines the chemical activity of selenium and its functional health effects eg towards cancers, in the heart and brain. It also covers other areas such as functional food enrichment, whole body metabolism, and the effects of selenium deficiency on health. Part of The Food and Nutritional Components in Focus series, this edited volume pools knowledge across scientific disciplines in a way that increases its applicability to a wide range of audiences. Victor Preedy’s own distinguished career in nutritional science has made him a prolific author of research articles and books in this area, and this project fills a gap in providing comprehensive synopses of food substances. Chemists, analytical scientists, forensic scientists, food scientists, as well as course lecturers will all benefit from this interdisciplinary title written by international experts in this area. |
| superdex 75: 4th Kuala Lumpur International Conference on Biomedical Engineering 2008 Noor Azuan Abu Osman, Prof. Ir. Dr Fatimah Ibrahim, Wan Abu Bakar Wan Abas, Herman Shah Abdul Rahman, Hua Nong Ting, 2008-07-30 It is with great pleasure that we present to you a collection of over 200 high quality technical papers from more than 10 countries that were presented at the Biomed 2008. The papers cover almost every aspect of Biomedical Engineering, from artificial intelligence to biomechanics, from medical informatics to tissue engineering. They also come from almost all parts of the globe, from America to Europe, from the Middle East to the Asia-Pacific. This set of papers presents to you the current research work being carried out in various disciplines of Biomedical En- neering, including new and innovative researches in emerging areas. As the organizers of Biomed 2008, we are very proud to be able to come-up with this publication. We owe the success to many individuals who worked very hard to achieve this: members of the Technical Committee, the Editors, and the Inter- tional Advisory Committee. We would like to take this opportunity to record our thanks and appreciation to each and every one of them. We are pretty sure that you will find many of the papers illuminating and useful for your own research and study. We hope that you will enjoy yourselves going through them as much as we had enjoyed compiling them into the proceedings. Assoc. Prof. Dr. Noor Azuan Abu Osman Chairperson, Organising Committee, Biomed 2008 |
| superdex 75: Comprehensive Sampling and Sample Preparation Josep M. Bayona, Paola Dugo, X. Chris Le, Hian Kee Lee, Xing-Fang Li, Heather Lord, 2012-12-31 Comprehensive Sampling and Sample Preparation is a complete treatment of the theory and methodology of sampling in all physical phases and the theory of sample preparation for all major extraction techniques. It is the perfect starting point for researchers and students to design and implement their experiments and support those experiments with quality-reviewed background information. In its four volumes, fundamentals of sampling and sample preparation are reinforced through broad and detailed sections dealing with Biological and Medical, Environmental and Forensic, and Food and Beverage applications. The contributions are organized to reflect the way in which analytical chemists approach a problem. It is intended for a broad audience of analytical chemists, both educators and practitioners of the art and can assist in the preparation of courses as well in the selection of sampling and sample preparation techniques to address the challenges at hand. Above all, it is designed to be helpful in learning more about these topics, as well as to encourage an interest in sampling and sample preparation by outlining the present practice of the technology and by indicating research opportunities. Sampling and Sample preparation is a large and well-defined field in Analytical Chemistry, relevant for many application areas such as medicine, environmental science, biochemistry, pharmacology, geology, and food science. This work covers all these aspects and will be extremely useful to researchers and students, who can use it as a starting point to design and implement their experiments and for quality-reviewed background information There are limited resources that Educators can use to effectively teach the fundamental aspects of modern sample preparation technology. Comprehensive Sampling and Sample Preparation addresses this need, but focuses on the common principles of new developments in extraction technologies rather than the differences between techniques thus facilitating a more thorough understanding Provides a complete overview of the field. Not only will help to save time, it will also help to make correct assessments and avoid costly mistakes in sampling in the process Sample and sample preparation are integral parts of the analytical process but are often less considered and sometimes even completely disregarded in the available literature. To fill this gap, leading scientists have contributed 130 chapters, organized in 4 volumes, covering all modern aspects of sampling and liquid, solid phase and membrane extractions, as well as the challenges associated with different types of matrices in relevant application areas |
| superdex 75: Precision Vaccinology for Infectious Diseases Mahbuba Rahman, Mehdi Adeli, Herb E. Schellhorn, Puthen Veettil Jithesh, Ofer Levy, 2024-04-29 The Human body is a vast network of interacting genes, proteins, and metabolites. These components, which may be considered host factors, change under disease, treatment or healthy condition. While treatment of many diseases depends on therapeutic drugs, vaccines remain the most effective long-term public health intervention to prevent infectious diseases. To date, vaccines have been developed to treat entire populations with little provision for predisposing individual host factor differences. However, the use and application of vaccines is facing multiple challenges with increasing numbers of vaccine non-responders and vaccine-relapsed individuals. The cause of this complication is partially due to host-factors. Another challenge is the adverse effects of vaccines in patients with primary immunodeficiency or autoimmune diseases, as well as vaccine-waning immunity in ageing populations, obese populations, or those with co-infection. To overcome these challenges, the solution may be the design, and formulation of precision vaccines, which are patient-specific. |
| superdex 75: Flavins and Flavoproteins 1993 Kunio Yagi, 2020-10-12 No detailed description available for Flavins and Flavoproteins 1993. |
| superdex 75: Chemical and Functional Properties of Food Proteins Zdzislaw E. Sikorski, 2001-06-22 Chemical and Functional Properties of Food Proteins presents the current state of knowledge on the content of proteins in food structures, the chemical, functional, and nutritive properties of food proteins, the chemical and biochemical modification of proteins in foods during storage and processing, and the mutagenicity and carcinogenicity of nitrogenous compounds. It emphasizes the structure-function relationship as well as the effects of practical conditions applied in food processing on the biochemical and chemical reactions in food proteins and food product quality. The first ten chapters discuss structure-function relationships, methods of analysis of nitrogenous compounds, chemical and enzymatic modifications, nutritive roles, and mutagenicity and carcinogenicity of food proteins. The following six chapters describe the proteins of meat and fish, milk, eggs, cereals, legumes, oilseeds and single cell organisms, and present detailed information on the effects of conditions applied in storage and processing on the reactions in proteins and their impact on quality attributes of food products. |
| superdex 75: Surface and Interfacial Aspects of Cell Adhesion Alain Carre, Kash L. Mittal, 2011-01-19 Cell adhesion comes into play in almost all domains of life. The range of situations in which it occurs, involving organisms, living tissues, microorganisms or single cells, is endless. Cell adhesion is involved in the binding of a cell to a surface, extracellular matrix, or another cell using cell adhesion molecules. It is crucial in the formation |
