Stable Isotope Labeling By Amino Acids In Cell Culture Silac
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Stable Isotope Labeling By Amino Acids In Cell Culture Silac
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Author : Bettina Warscheid
language : en
Publisher:
Release Date : 2014
Stable Isotope Labeling By Amino Acids In Cell Culture Silac written by Bettina Warscheid and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2014 with Amino acids categories.
Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols provides a synopsis of a large array of different SILAC methods by presenting a set of protocols that have been established by renowned scientists and their working groups.These include methods and protocols for the labeling of various model organisms as well as advanced strategies relying on SILAC, e.g. for the analysis of protein interactions, the mapping of posttranslational modifications or the characterization of subcellular proteomes. 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-bystep, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols will serve students and experienced scientists alike as a valuable reference of how to make use of the SILAC technology for their own research.
Stable Isotope Labeling By Amino Acids In Cell Culture Silac
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Author : Shao-En Ong
language : en
Publisher:
Release Date : 2004
Stable Isotope Labeling By Amino Acids In Cell Culture Silac written by Shao-En Ong and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2004 with categories.
Silac
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Author : Jose L. Luque-Garcia
language : en
Publisher: Springer Nature
Release Date : 2022-11-12
Silac written by Jose L. Luque-Garcia and has been published by Springer Nature this book supported file pdf, txt, epub, kindle and other format this book has been release on 2022-11-12 with Science categories.
This detailed volume explores the most recent methodologies and applications of stable isotope labeling by amino acids in cell culture (SILAC). With the development of new technologies in the field of mass spectrometry and bioinformatics, SILAC-based methods have become the first tool of choice in quantitative proteomics either on its own or in combination with other more recently developed approaches, which is elucidated in the chapters of this book. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, SILAC: Methods and Protocols serves as an ideal guide for researchers working in the area of functional proteomics and other aspects of protein science.
The Use Of Stable Isotope Labeling With Amino Acids In Cell Culture Silac To Study Ivermectin Mediated Molecular Pathway Changes In Human Ovarian Cancer Cells
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Author : Xianquan Zhan
language : en
Publisher:
Release Date : 2019
The Use Of Stable Isotope Labeling With Amino Acids In Cell Culture Silac To Study Ivermectin Mediated Molecular Pathway Changes In Human Ovarian Cancer Cells written by Xianquan Zhan and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019 with Science categories.
Stable isotope labeling with amino acids in cell culture (SILAC) was to use isotopic essential amino acids to replace the original amino acids for cell culture and passage for 8,Äì10 generations, followed by mass spectrometry to identify proteins and the isotopic abundance difference to quantify proteins. SILAC can be used to characterize proteomic changes, and analyze protein turnover, protein interactions, and dynamic changes with quantitative accuracy, and high reproducibility. For this study, SILAC ,Äúlight,Äù (L-Lysine-2HCl [12C6, 14N2], L-Arginine-HCl [12C6, 14N4])- or ,Äúheavy,Äù (L-Lysine-2HCl [13C6, 15N2], L-Arginine-HCl [13C6, 15N4])-labeling RPMI 1640 medium was used to culture human ovarian cancer TOV-21G cells for 10 passages, followed by the treatment of 0.1% dimethylsulfoxide for 24 h and 20 ¬μM ivermectin for 24¬†h, respectively. The light- and heavy-isotope-labeled proteins were equally mixed (1:1) for digestion with trypsin. The tryptic peptide mixture was fractionated with liquid chromatography and analyzed with tandem mass spectrometry. In total, 4,447 proteins were identified in ivermectin-treated TOV-21G cells in relation to controls. Those proteins were enriched in 89 statistically significant signaling pathways and 62 statistically significant biological processes. These findings clearly demonstrated that SILAC quantitative proteomics was a useful and reliable method to study ivermectin-related proteomic changes in cancer cells, which in combination with molecular pathway networks and biological processes enrichments provided more comprehensive insights into molecular mechanisms of ivermectin in inhibiting TOV-21G cells.
Stable Isotope Labeling By Amino Acids In Cell Culture Silac
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Author : Bettina Warscheid
language : en
Publisher: Humana Press
Release Date : 2014-07-25
Stable Isotope Labeling By Amino Acids In Cell Culture Silac written by Bettina Warscheid and has been published by Humana Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2014-07-25 with Science categories.
Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols provides a synopsis of a large array of different SILAC methods by presenting a set of protocols that have been established by renowned scientists and their working groups. These include methods and protocols for the labeling of various model organisms as well as advanced strategies relying on SILAC, e.g. for the analysis of protein interactions, the mapping of posttranslational modifications or the characterization of subcellular proteomes. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC): Methods and Protocols will serve students and experienced scientists alike as a valuable reference of how to make use of the SILAC technology for their own research.
Advancements Of Mass Spectrometry In Biomedical Research
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Author : Alisa G. Woods
language : en
Publisher: Springer
Release Date : 2016-09-24
Advancements Of Mass Spectrometry In Biomedical Research written by Alisa G. Woods and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2016-09-24 with Science categories.
This volume explores the use of mass spectrometry for biomedical applications. Chapters focus on specific therapeutic areas such as oncology, infectious disease and psychiatry. Additional chapters focus on methodology as well as new technologies and instrumentation. This volume provides readers with a comprehensive and informative manual that will allow them to appreciate mass spectrometry and proteomic research but also to initiate and improve their own work. Thus the book acts as a technical guide but also a conceptual guide to the newest information in this exciting field. Mass spectrometry is the central tool used in proteomic research today and is rapidly becoming indispensable to the biomedical scientist. With the completion of the human genome project and the genomic revolution, the proteomic revolution has followed closely behind. Understanding the human proteome has become critical to basic and clinical biomedical research and holds the promise of providing comprehensive understanding of human physiological processes. In addition, proteomics and mass spectrometry are bringing unprecedented biomarker discovery and are helping to personalize medicine.
Quantitative Proteomics By Mass Spectrometry
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Author : Salvatore Sechi
language : en
Publisher: Methods in Molecular Biology
Release Date : 2018-04-15
Quantitative Proteomics By Mass Spectrometry written by Salvatore Sechi and has been published by Methods in Molecular Biology this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018-04-15 with Science categories.
This volume describes prominent methodologies developed by laboratories that have been leading the field of quantitative proteomics by mass spectrometry. The procedures for performing the experiments are described in an easy-to-understand manner with many technical details that usually are not reported in typical research articles. This second edition of Quantitative Proteomics by Mass Spectrometry provides a broad perspective of the methodologies used for quantifying proteins and post-translational modifications in different types of biomedical specimens. 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 thorough, Quantitative Proteomics by Mass Spectrometry, Second Edition is a valuable resource to help researchers understand and learn about the latest tools used in the study of quantitative proteomics by mass spectrometry.
Synthesis Of Violacein For Stable Isotopic Labeling By Amino Acids In Cell Culture
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Author : Awura Ama A Oppong-Holmes
language : en
Publisher:
Release Date : 2018
Synthesis Of Violacein For Stable Isotopic Labeling By Amino Acids In Cell Culture written by Awura Ama A Oppong-Holmes and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018 with Antibiotics categories.
Violacein is an intensely purple microbial pigment with an arsenal of biological activity. A structural analog to the better-known bis-indole alkaloids staurosporine and rebeccamycin, violacein has been touted as an antimicrobial and anticancer therapeutic. It is most frequently obtained as a biosynthetic secondary metabolite from bacterial cell cultures. However, we have developed an efficient, environmentally benign methodology for synthesizing the natural product via a 3-component condensation in polyethylene glycol. Violacein, along with a series of analogs have been prepared in our laboratory. The deoxyviolacein analog has been investigated for its effects against cancer cell migration in HeLa. This initial screening has served as the impetus to study deoxyviolacein's specific interactions in cancer cells via proteomics. In this project, the synthesis of the deoxyviolacein scaffold for solid support is designed, in preparation for SILAC (Stable Isotope Labeling by Amino acids in Cell culture) analysis. The synthesis requires preparation of 5-hydroxymethyl-1H-indole, reported here. The main objective of this project is to design and synthesize a functional analog that can be used to determine the protein interactions responsible for the biological activity of violacein.
Quantitative Methods In Proteomics
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Author : Katrin Marcus
language : en
Publisher: Humana Press
Release Date : 2012-06-08
Quantitative Methods In Proteomics written by Katrin Marcus and has been published by Humana Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-06-08 with Science categories.
Protein modifications and changes made to them, as well as the quantities of expressed proteins, can define the various functional stages of the cell. Accordingly, perturbations can lead to various diseases and disorders. As a result, it has become paramount to be able to detect and monitor post-translational modifications and to measure the abundance of proteins within the cell with extreme sensitivity. While protein identification is an almost routine requirement nowadays, reliable techniques for quantifying unmodified proteins (including those that escape detection under standard conditions, such as protein isoforms and membrane proteins) is not routine. Quantitative Methods in Proteomics gives a detailed survey of topics and methods on the principles underlying modern protein analysis, from statistical issues when planning proteomics experiments, to gel-based and mass spectrometry-based applications. The quantification of post-translational modifications is also addressed, followed by the “hot” topics of software and data analysis, as well as various overview chapters which provide a comprehensive overview of existing methods in quantitative proteomics. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Quantitative Methods in Proteomics serves as a comprehensive and competent overview of the important and still growing field of quantitative proteomics.
Improvements To Stable Isotope And Label Free Quantitative Proteomic Methods
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Author :
language : en
Publisher:
Release Date : 2013
Improvements To Stable Isotope And Label Free Quantitative Proteomic Methods written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2013 with categories.
The following thesis presents strategies for improving throughput and accuracy of mass spectrometry based proteomic methods and the application of these methods to study biological systems. A brief description of qualitative and quantitative proteomics is presented in Chapter 1. In Chapter 2, I report the first description of using mass defects for stable isotope labeling with amino acids in cell culture (SILAC). Using amino acid isotopologues with sub dalton mass differences permits increased multiplexing without concomitant increase in spectral complexity that occurs with SILAC based methods. Chapter 3 introduces quantitative amine reactive mass defect labels. These labels offer quantitation similar to mass defect SILAC but are compatible with tissue sample analysis. Chapter 4 describes a gas phase purification method that improves the accuracy and dynamic range of isobaric labeling by reducing measurement interference derived from precursor co-isolatation. Gas phase purification is accomplished by reacting selected peptide precursors with a proton transfer reagent, followed by re-isolation, fragmentation, and analysis of the charge reduced peptide. This method is then applied to measure changes in the mouse liver mitochondrial acetylome in Chapters 5 and 6. Dramatic changes associated with SIRT3 knockout and calorie restriction are described in Chapter 5. More subtle effects of obesity and fasting are presented in Chapter 6. Finally, a high-throughput workflow to comprehensively sequence a yeast proteome is reported in Chapter 7. This workflow offers a path toward high-throughput label-free proteomics experiments.