Capillary Electrophoresis with Post-column Laser-induced Fluorescence Detection of Proteins

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ISBN 13 :
Total Pages : 42 pages
Book Rating : 4.:/5 (932 download)

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Book Synopsis Capillary Electrophoresis with Post-column Laser-induced Fluorescence Detection of Proteins by :

Download or read book Capillary Electrophoresis with Post-column Laser-induced Fluorescence Detection of Proteins written by and published by . This book was released on 1996 with total page 42 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary electrophoresis with laser induced fluorescence detection (CE-LIF) is one of the alternative methods being developed for fast and sensitive assays of proteins and peptides. This report summarizes CE-LIF instrument development, efforts at labelling proteins, tests conducted to prove the applicability of CE-LIF to the analysis of samples, and results of a comparison of CE-LIF with some other techniques. Appendices include an instrument operating procedure and a paper that summarizes the experimental details of the protein labelling procedure and a CE-LIF technique for sub-nanomolar assay of proteins.

A New Post-column Reactor-laser Induced Fluorescence Detector for Capillary Electrophoresis

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Publisher :
ISBN 13 :
Total Pages : 43 pages
Book Rating : 4.:/5 (683 download)

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Book Synopsis A New Post-column Reactor-laser Induced Fluorescence Detector for Capillary Electrophoresis by :

Download or read book A New Post-column Reactor-laser Induced Fluorescence Detector for Capillary Electrophoresis written by and published by . This book was released on 1996 with total page 43 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary zone electrophoresis (CZE), a powerful separation method based on the differential migration of charged species under the influence of an electric field, has been widely used for separations covering from small ions to big biomolecules. Chapter 1 describes the method, then discusses detection of the separated analytes by laser induced fluorescence and by chemical derivatization, and the use of O-phthaldialdehyde (OPA) as a post-column reagent. Chapter 2 describes a post-column reactor which uses two narrow bore capillaries connected coaxially. This reactor differs from other coaxial reactors in terms of capillary dimensions, reagent flow control, ease of construction and most importantly, better limits of detection. The derivatization reagent is electroosmotically driven into the reaction capillary and the reagent flow rate is independently controlled by a high voltage power supply. Amino acids, amines and proteins, derivatized by OPA/2-mercaptoethanol using this post-column reactor coupled with LIF detection, show low attomole mass limits of detection, and for the first time, the authors demonstrate single cell capability with a post-column derivatization scheme. The single cell capability shows that this reactor could find applications in assaying non-fluorescent or electrochemically inactive components in individual biological cells in the future.

Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection

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ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (11 download)

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Book Synopsis Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection by : Michael D. Harvey

Download or read book Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection written by Michael D. Harvey and published by . This book was released on 2001 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The trend towards high throughput applications and miniaturization necessitates approaches capable of microlitre volume sampling and low protein concentration detection. Furthermore, one of the major trends in high throughput screening is the growing replacement of technologies that depend on radioactivity to generate a signal with those that rely on fluorescence. This trend towards non-radioactive detection in general can be understood by some of the advantages inherent to these methods over radioactive modes. These include a significant reduction in safety concerns leading to a relaxation of strict laboratory procedures, elimination of expensive waste disposal, extended shelf-life of labeled reagents, and the possibility of acquiring multiplexed data through the spectral isolation of different wavelength signals. A variety of capillary electrophoretic (CE) approaches utilizing laser-induced fluorescence (LIF) have thus been developed, providing researchers with valuable tools in protein analysis. Various covalent and non-covalent fluorescent derivatization approaches have been investigated, with emphasis on biochemical and/or clinical applications. The non-covalent dye, NanoOrange, is used as a clinical diagnostic tool for early disease diagnosis, quantitating nanomolar concentrations of human serum albumin in solution, and obtaining fluorescence-based biofluid profiles. An alternate non-covalent labeling approach utilizing the fluorescent probe, Sypro Red, and capillary gel electrophoresis allows for rapid, sensitive analysis of protein sample purity as well as molecular weight determination. These two non-covalent approaches are complemented by the development of a fluorescent Insulin-Like Growth Factor-I (IGF-I) analog for use in bioanalytical applications. Specific derivatization reaction conditions were developed to selectively label the N-terminus of the analog hence preserve biological activity. High-performance liquid chromatography and electrospray mass spectrometry were used to confirm the extent of labeling and modification site. Antibody recognition of this fluorescent analog was evaluated using CE-LIF, illustrating the clinical utility of this diagnostic reagent. In addition to the above CE-LIF approaches, a fourth capillary electrophoretic tool is provided for the clinical chemist. Rapid analysis of biofluids is of significant importance in early disease diagnosis. As such, an extensive CE-based analysis of human seminal plasma is presented. Separation conditions, sample stability, and protein/non-protein zone identification issues are addressed. This study and the CE-LIF methodologies discussed above represent original approaches to nanoscale protein analysis.

Capillary Electrophoresis of Proteins and Peptides

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Publisher : Springer Science & Business Media
ISBN 13 : 159259798X
Total Pages : 340 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Capillary Electrophoresis of Proteins and Peptides by : Mark A. Strege

Download or read book Capillary Electrophoresis of Proteins and Peptides written by Mark A. Strege and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 340 pages. Available in PDF, EPUB and Kindle. Book excerpt: Throughout the more than 20 years that have followed the beginnings of capillary electrophoresis (CE), its application to the analysis of proteins and peptides has continued to be reliable, versatile, and productive. Over time, CE has matured to become a superb complement to HLPC, and in many cases has also evolved as an automated and quantitative replacement for conventional slab gel electrophoresis methods such as SDS-PAGE and isoelectric focusing. Within Capillary Electrophoresis of Proteins and Peptides, we have assembled contributions from researchers who are applying state-of-the-art CE for protein and peptide analysis, including topics that we believe are of great potential both in the present and for the future. In comparison to traditional separation methods, CE represents a miniaturized analysis technique (especially in its microchip-based format) that is highly dependent upon the basic fundamentals of effective sample recovery and high sensitivity detection. With these issues in mind, Chapters 1–4 describe recently developed approaches for both capillary coatings and analyte detection via laser-induced fluorescence. Since the discipline of biotechnology has established itself as a primary platform for the application of CE to the analysis of proteins and peptides, Chapters 5–7 demonstrate a variety of examples of the specific techniques that have been applied for the development of biopharmaceuticals and their commercialization. The methods covered here include also the analysis of oligosaccharides from glycoproteins.

Handbook of Capillary Electrophoresis Applications

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Publisher : Springer Science & Business Media
ISBN 13 : 9400915616
Total Pages : 750 pages
Book Rating : 4.4/5 (9 download)

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Book Synopsis Handbook of Capillary Electrophoresis Applications by : H. Shintani

Download or read book Handbook of Capillary Electrophoresis Applications written by H. Shintani and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 750 pages. Available in PDF, EPUB and Kindle. Book excerpt: Over the last decade, high performance Capillary electrophoresis (HPCE) has emerged as a powerful and versatile separation technique that promises to rival high performance liquid chromatography when applied to the separation of both charged and neutral species. The high speed and high separation efficiency which can be attained using any of the various modes of HPCE has resulted in the increased use of the technique in a range of analytical environments. The procedures are, however, still in the early stages of development and several barriers remain to their adoption as the technique of choice for a range of analytical problems. One such barrier is the selection and optimization of the conditions required to achieve reproducible separations of analytes and it is in this area that this new book seeks to give assistance. The book is written by an international team of authors, drawn from both academic and industrial users, and the manufacturers of instruments. At its heart are a number of tables, divided into specific application areas. These give details of published separations of a wide range of archetypal analytes, the successful separation conditions and the matrix in which they were presented. These tables are based on separations reported since 1992 and are fully referenced to the original literature. The tables are supported by discussions of the problems that a particular area presents and the strategies and solutions adopted to overcome them. The general areas covered are biochemistry, pharmaceutical science, bioscience, ion analysis, food analysis and environmental science.

Two-wavelength Laser Induced Fluorescence Detection for Capillary Electrophoresis of Proteins and Peptides

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Publisher :
ISBN 13 :
Total Pages : 136 pages
Book Rating : 4.:/5 (42 download)

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Book Synopsis Two-wavelength Laser Induced Fluorescence Detection for Capillary Electrophoresis of Proteins and Peptides by : Guido Fridolin Verbeck

Download or read book Two-wavelength Laser Induced Fluorescence Detection for Capillary Electrophoresis of Proteins and Peptides written by Guido Fridolin Verbeck and published by . This book was released on 1999 with total page 136 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Protein Analysis by Capillary Electrophoresis with Laser-induced Fluorescence Detection

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Publisher :
ISBN 13 :
Total Pages : 290 pages
Book Rating : 4.:/5 (584 download)

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Book Synopsis Protein Analysis by Capillary Electrophoresis with Laser-induced Fluorescence Detection by : Franklin H. Welder

Download or read book Protein Analysis by Capillary Electrophoresis with Laser-induced Fluorescence Detection written by Franklin H. Welder and published by . This book was released on 2004 with total page 290 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Clinical and Forensic Applications of Capillary Electrophoresis

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Publisher : Springer Science & Business Media
ISBN 13 : 1592591205
Total Pages : 449 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Clinical and Forensic Applications of Capillary Electrophoresis by : John R. Petersen

Download or read book Clinical and Forensic Applications of Capillary Electrophoresis written by John R. Petersen and published by Springer Science & Business Media. This book was released on 2001-04-20 with total page 449 pages. Available in PDF, EPUB and Kindle. Book excerpt: Because capillary electrophoresis (CE) is driving the automation of manual and semi-manual techniques, conserves precious samples, and minimizes the use of hazardous organic chemicals, it constitutes a powerful new methodology for the clinical laboratory. In Clinical and Forensic Applications of Capillary Electrophoresis, John R. Petersen and Amin A. Mohammad, along with a panel of leading basic and clinical investigators, review those CE methods that are now replacing many routine serum and blood tests in clinical and forensic laboratories. Major areas reviewed include the coating of columns; the analysis of serum, urine, and CSF proteins and paraproteins; abnormal hemoglobins and hemoglobin Alc; peptides, amino and organic acids; therapeutic drugs; drugs of abuse; viral load; and short tandem repeats (STR). The methods discussed include capillary zone, micellar, electrokinetic, capillary gel, and nonaqueous electrophoresis. The authors show how a variety of detection methods (UV-visible, fluorescence, and MS) can be used in conjunction with CE to detect a broad array of analytes, ranging from serum proteins to PCR products to drugs of forensic importance. Innovative and highly practical, Clinical and Forensic Applications of Capillary Electrophoresis demonstrates the power and versatility of CE-not only to develop new assays, but also to markedly simplify today's clinical and forensic laboratory methodology.

Analysis of Large Biomolecules by Capillary Electrophoresis with Laser-induced Fluorescence Detection

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ISBN 13 :
Total Pages : 326 pages
Book Rating : 4.:/5 (584 download)

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Book Synopsis Analysis of Large Biomolecules by Capillary Electrophoresis with Laser-induced Fluorescence Detection by : Pertti Juha Viskari

Download or read book Analysis of Large Biomolecules by Capillary Electrophoresis with Laser-induced Fluorescence Detection written by Pertti Juha Viskari and published by . This book was released on 2003 with total page 326 pages. Available in PDF, EPUB and Kindle. Book excerpt:

An Investigation on Capillary Electrophoresis-Based Immunoassay with Laser Induced Fluorescence Detection

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Publisher :
ISBN 13 : 9781374748651
Total Pages : pages
Book Rating : 4.7/5 (486 download)

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Book Synopsis An Investigation on Capillary Electrophoresis-Based Immunoassay with Laser Induced Fluorescence Detection by : Jianping Ou

Download or read book An Investigation on Capillary Electrophoresis-Based Immunoassay with Laser Induced Fluorescence Detection written by Jianping Ou and published by . This book was released on 2017-01-27 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Capillary Electrophoresis

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Publisher : Springer Science & Business Media
ISBN 13 : 1588295397
Total Pages : 808 pages
Book Rating : 4.5/5 (882 download)

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Book Synopsis Capillary Electrophoresis by : Philippe Schmitt-Kopplin

Download or read book Capillary Electrophoresis written by Philippe Schmitt-Kopplin and published by Springer Science & Business Media. This book was released on 2008-02-25 with total page 808 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book presents a selection of current capillary electrophoresis methods used to separate representative types of molecules and particles and in combination with different detection techniques. It includes practical details which are hard to find elsewhere. The volume is intended for beginners in the field and provides an overview of the technique and a starting point for the exploration of the defined literature on different application topics.

Analysis by Capillary Electrophoresis with Laser-induced Fluorescence Detection

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Publisher :
ISBN 13 :
Total Pages : 289 pages
Book Rating : 4.:/5 (794 download)

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Book Synopsis Analysis by Capillary Electrophoresis with Laser-induced Fluorescence Detection by : Tara Lee Massie

Download or read book Analysis by Capillary Electrophoresis with Laser-induced Fluorescence Detection written by Tara Lee Massie and published by . This book was released on 2010 with total page 289 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) is a versatile and sensitive analytical tool with many potential applications. Its advantages over other chromatographic methods include minimal solvent and sample requirements, low waste volumes, simple instrumentation, and diversity of analytes studied. CE is one of the most efficient separation techniques available for the analysis of both large and small molecules, but because few analytes are natively fluorescent, it is necessary to bind them with fluorescent tags if LIF detection is to be employed. These tags increase sensitivity and provide additional separation from components of the sample that do not bind the tags. In this dissertation, four applications of CE-LIF for the determination of tagged analytes are documented. First, a rapid CE-LIF method was developed for the determination of gamma-aminobutyric acid (GABA) and other important brain biogenic amines and amino acids present in biological samples at low levels. The high efficiency and sensitivity necessary to identify toxicologically relevant changes in neurotransmitter levels in brain tissue and microdialysates following Mn exposure was achieved, and the method required minimum sample work-up and quantities of sample for analysis. A linear relationship between the concentration of GABA and the CE-LIF peak height was established, with correlation coefficients of 0.989 or better for four neurotransmitters. Second, the use of two squarylium-based fluorescent dyes (bis-SQHN-4d and SQHN-3c) as noncovalent protein labels was investigated. The fluorescence emissions of both dyes were enhanced upon complexation with model proteins, demonstrating promise as noncovalent fluorescent protein labels. Third, the nature of interactions between two boronic acid-based probes (SQTM-BA1 and ANQW-BA1) and two model proteins (bovine serum albumin (BSA) and hemagglutinin (HA)) have been explored. The complex between SQTM-BA1 and BSA was the only complex to show increased fluorescence emission, and the role of electrostatic interactions between SQTM-BA1 and BSA in complex formation was explored. Lastly, CE-LIF methods were developed to label and quantify turnip yellow mosaic virus (TYMV). Noncovalent labels Red-1c and NN127 effectively labeled the surface proteins of the virus, and the labeled viruses were then analyzed by CE-LIF. The CE-LIF methods developed herein for the analysis of neurotransmitters, free solution proteins, and viruses demonstrate the versatility of the technique, especially when coupled with fluorescent labeling.

Handbook of Capillary Electrophoresis, Second Edition

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Publisher : CRC Press
ISBN 13 : 9780849324987
Total Pages : 918 pages
Book Rating : 4.3/5 (249 download)

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Book Synopsis Handbook of Capillary Electrophoresis, Second Edition by : James P. Landers

Download or read book Handbook of Capillary Electrophoresis, Second Edition written by James P. Landers and published by CRC Press. This book was released on 1996-12-23 with total page 918 pages. Available in PDF, EPUB and Kindle. Book excerpt: Because new information was discovered at an incredible rate since the publication of the successful first edition of this Handbook, this fully updated second edition covers all areas of interest in the field of capillary electrophoresis (CE). A relatively new technology, CE is a principle method for studying the physicochemical properties of proteins, peptides, and other macromolecules. Where applicable, the 30 chapters provide basic underlying theories as well as application-oriented aspects of each technique.Keep up with all the developments in this growing field with the Handbook of Capillary Electrophoresis, Second Edition - a complete guide to the fundamentals of CE and the latest research. The chapters are organized into five units: Modes: Presents a theoretical development of the basic principles governing separation with several modes, including CEC, and discusses their practical aspects. Analyte: Applies CE to the analysis of a specific class of analytes, including organic and inorganic ions, pharmaceuticals, glycoconjugates, peptides, proteins, and DNA fragments. Fundamental Aspects of CE: Technique-oriented information for the practitioner, including the importance of the sample matrix, on-line preconcentration of samples, modes of detection, and specific aspects of CE data analysis. Applications of CE: Includes single cell analysis, CE in DNA sequencing, CE as a clinical diagnostic tool, identifying and quantifying drugs, and for characterizing interacting species. Specialized Aspects of CE: Discusses interfacing CE with mass spectrometry, high-volume throughput continuous CE, microchip CE, control of EOF, and much more. The Handbook of Capillary Electrophoresis, Second Edition, pulls together diverse areas and applications of CE, resulting in an excellent tool for scientists involved in biotechnology and clinical chemistry, as well as the pharmaceutical, bioscience, chemical, and instrument-manufacturing industries. With an applications-oriented focus, the handbook is also a superb manual for workshops, seminars, and graduate courses in separation science.

Capillary Gel Electrophoresis

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Publisher : Newnes
ISBN 13 : 0080931359
Total Pages : 391 pages
Book Rating : 4.0/5 (89 download)

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Book Synopsis Capillary Gel Electrophoresis by : Andras Guttman

Download or read book Capillary Gel Electrophoresis written by Andras Guttman and published by Newnes. This book was released on 2021-12-04 with total page 391 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent overview of the key application areas of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods. It not only gives readers a better understanding of how to utilize this technology, but also provides insights into how to determine which method will provide the best technical solutions to particular problems. This book can also serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses. Covers all theoretical and practical aspects of capillary gel electrophoresis Excellent overview of the key applications of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods Teaches readers how to use the technology and select methods that are ideal for fundamental problems Can serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses

SDS-capillary Gel Electrophoresis of Derivatized Proteins Using Laser-induced Fluorescence Detection

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Publisher :
ISBN 13 :
Total Pages : 142 pages
Book Rating : 4.:/5 (721 download)

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Book Synopsis SDS-capillary Gel Electrophoresis of Derivatized Proteins Using Laser-induced Fluorescence Detection by : Eric Thomas Wise

Download or read book SDS-capillary Gel Electrophoresis of Derivatized Proteins Using Laser-induced Fluorescence Detection written by Eric Thomas Wise and published by . This book was released on 1995 with total page 142 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Practical Capillary Electrophoresis

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Publisher : Elsevier
ISBN 13 : 0080539343
Total Pages : 425 pages
Book Rating : 4.0/5 (85 download)

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Book Synopsis Practical Capillary Electrophoresis by : Robert Weinberger

Download or read book Practical Capillary Electrophoresis written by Robert Weinberger and published by Elsevier. This book was released on 2000-04-18 with total page 425 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the 1980s, capillary electrophoresis (CE) joined high-performance liquid chromatography (HPLC) as the most powerful separation technique available to analytical chemists and biochemists. Published research using CE grew from 48 papers in the year of commercial introduction (1988) to 1200 in 1997. While only a dozen major pharmaceutical and biotech companies have reduced CE to routine practice, the applications market is showing real or potential growth in key areas, particularly in the DNA marketplace for genomic mapping and forensic identification. For drug development involving small molecules (including chiral separations), one CE instrument can replace 10 liquid chromatographs in terms of speed of analysis. CE also uses aqueous rather than organic solvents and is thus environmentally friendlier than HPLC. The second edition of Practical Capillary Electrophoresis has been extensively reorganized and rewritten to reflect modern usage in the field, with an emphasis on commercially available apparatus and reagents. This authoritative and very comprehensible treatment builds on the author's extensive experience as an instructor of short courses for the American Chemical Society and for industry. Illustrated with detailed diagrams of electrophoretic phenomena Offers step-by-step methods development schemes Presents techniques for developing quantitative, robust, and precise methods Includes an extensive troubleshooting guide Updates and greatly expands on the first edition-more than 50% of the text is new Written by an internationally recognized scientist who is an instructor for American Chemical Society short courses on HPCE

Development of High Sensitivity Capillary Electrophoresis Fluorescence System for the Detection of Biological Molecules:

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Publisher :
ISBN 13 :
Total Pages : 31 pages
Book Rating : 4.:/5 (933 download)

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Book Synopsis Development of High Sensitivity Capillary Electrophoresis Fluorescence System for the Detection of Biological Molecules: by :

Download or read book Development of High Sensitivity Capillary Electrophoresis Fluorescence System for the Detection of Biological Molecules: written by and published by . This book was released on 2000 with total page 31 pages. Available in PDF, EPUB and Kindle. Book excerpt: One class of chemical & biological weapons consists of toxic proteins & peptides produced by biological organisms, or mid-spectrum agents. The research described in this report considered the analysis & detection of these mid-spectrum agents based on chemical methods, specifically via the use of capillary electrophoresis (CE) coupled with laser induced fluorescence (LIF). The report first presents a brief literature review and then highlights results of three research themes. Research papers detailing the experiments & results of these themes are appended. The first paper reports a method for assay of proteins at picomolar concentrations by CE, precolumn labelling, sub-micellar separation, and LIF detection. The second describes the use of sieving media for the separation of labeled proteins. The last paper addresses on-column labeling & analysis of proteins with a fluorogenic reagent. Overall results of the methods developed are compared to those achieved with ultraviolet absorbance detection.