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:
Download or read book Analysis of Biomolecules Using Interfacial Capillary Electrophoresis with Laser Induced Fluorescence Detection written by Lorna Alice Hunter Pehl and published by . This book was released on 2000 with total page 178 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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.
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.
Download or read book Applications of Capillary Electrophoresis and Laser induced Fluorescence Detection to the Analysis of Trace Species written by and published by . This book was released on 1995 with total page 52 pages. Available in PDF, EPUB and Kindle. Book excerpt: This Ph. D. Thesis describes several separation and detection schemes for the analysis of small volume and amount of samples, such as intracellular components and single enzymes developed during research. Indirect Laser-induced fluorescence detection and capillary electrophoresis were used to quantify lactate and pyruvate in single red blood cells. The assay of specific enzyme activities was achieved by monitoring the highly fluorescent enzymatic reaction product, NADH. LDH activity was found not to be a unique marker for diagnosis of leukemia. Reactions of single LDH-1 molecules were investigated by monitoring the reaction product with LIF detection.
Download or read book Applications of Capillary Electrophoresis and Laser induced Fluorescence Detection to the Analysis of Trace Species written by Qifeng Xue and published by . This book was released on 1995 with total page 258 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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:
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.
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.
Download or read book Investigations Into the Use of Capillary Electrophoresis for the Analysis of Nuclear Preparations and the Nuclear Localization of Fusion Proteins at the Single Cell Level written by Nilhan Gunasekera and published by . This book was released on 2003 with total page 328 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Capillary Zone Electrophoresis with Laser induced Fluorescence and Chemiluminescence Detection for Biomolecule Analysis written by Jian Ying Zhao and published by . This book was released on 1994 with total page 380 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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.
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.
Download or read book Capillary Electrophoresis in the Life Sciences written by Ante M. Krstulović and published by Elsevier Science & Technology. This book was released on 1997 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: High-performance capillary electrophoresis (CE) is at present widely considered as the most important milestone in the field of separation science of the last decade. The current developments in modern biology and medicine necessitate high-resolution microtechniques for the analysis of large biological molecules. Due to its high resolving power, high speed of analysis and low cost operation, CE has been extensively used in the analysis of proteins, peptides, oligonucleotides, DNA, DNA sequencing of genomic material, double-stranded DNA fragments in mutational analysis, restriction fragment mapping, etc. Its hybrids with liquid chromatography, such as micellar electrokinetic capillary chromatography and capillary electrochromatography have extended the use to relatively small organic molecules encountered in pharmaceutical, environmental, forensic and clinical analysis, as well as combinatorial chemistry and fundamental research, where its role has been complimentary to that of the well established HPLC techniques. Because of the dramatic decrease in sample volume (nanoliters and picoliters) even the analysis of single cell contents has been made possible. In order to overcome the problem of low sensitivity, on-line preconcentration techniques (isotachophoretic) sample stacking, electrochromatographic, affinity, microchips, etc.), as well as laser-induced fluorescence and various mass spectrometric techniques have been used. While the use of CE in the area of biomolecular separations is now well-established, its usefulness as an orthogonal technique with respect to HPLC in the analysis of small molecules is yet to be developed. It is hoped that the selected papers of this volume will illustrate further the tremendous potential of CE in the life sciences.
Download or read book Applications of Laser Induced Fluorescence Detection in Biomedical Analysis written by John Emerson Kuo and published by . This book was released on 1985 with total page 336 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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
Download or read book Detection Advances for Analysis of Biological Microenvironments Using Capillary Electrophoresis written by Jason Ben Shear and published by . This book was released on 1994 with total page 356 pages. Available in PDF, EPUB and Kindle. Book excerpt:
