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Capillary Electrophoretic Enzyme Assays

In the past years, capillary electrophoresis has become a frequently used technique for enzyme assays due to the high separation efficiency and versatility as well as small sample size and low consumption of chemicals. The capillary electrophoresis assays can be divided into two general c ...

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Capillary Electrophoresis for Protein Profiling of the Dimorphic, Pathogenic Fungus, Penicillium marneffei

Penicillium marneffei is an endemic, dimorphic fungus that exhibits very significant morbidity among immune compromised persons living or having traveled in Southeast Asia. The dimorphic nature of P. marneffei, which is believed to be a major contributing factor to infection by this fu ...

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Fluorescent Lipids as Probes for Sphingosine Kinase Activity by Capillary Electrophoresis

Capillary electrophoresis (CE) is one among a number of highly sensitive chemical separation techniques used to characterize single or a small number of cells and to develop assays of enzymatic activity. Other commonly used techniques include mass spectrometry and electrochemist ...

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Capillary ElectrophoresisMass Spectrometry of Carbohydrates

The development of methods for capillary electrophoresis (CE) with on-line mass spectrometric detection (CE/MS) is driven by the need for accurate, robust, and sensitive glycomics analysis for basic biomedicine, biomarker discovery, and analysis of recombinant protein therape ...

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Assessment of DNA Damage by Micellar Electrokinetic Chromatography

A simple and inexpensive MEKC method, which is able to assess base damage within DNA samples, is illustrated. After heat-acid hydrolysis of the DNA samples, both the percentage of each canonical DNA base and the relative amount of uncanonical DNA bases can be measured. This method is useful for an eval ...

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Approaches to Enhancing the Sensitivity of Carbohydrate Separations in Capillary Electrophoresis

Electrophoresis in both capillaries (CE) and microchips (ME) is an extremely powerful liquid phase-separation technique that is indispensable for the separation of carbohydrates. It is capable of separating both small mono- and disaccharides, through to more complex oligo- and pol ...

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Determination of Monosaccharides Derivatized with 2-Aminobenzoic Acid by Capillary Electrophoresis

Reducing monosaccharides were derivatized with 2-aminobenzoic acid (2-AA) through reductive amination using sodium cyanoborohydride as a reductant, and the derivatives were separated by capillary zone electrophoresis with UV detection using 50 mM sodium phosphate (pH 5.5) or 1 ...

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Determination of Mono-, Di-, and Oligosaccharides by Capillary Electrophoresis with Capacitively Coupled Contactless Conductivity Detection

Saccharides and chitooligosaccharides can be separated in electrophoretic conditions by raising the pH of the medium, which renders the corresponding alcoholate forms. These anionic species can be separated and detected with capacitively coupled contactless conductivity ...

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Capillary Electrophoresis for the Analysis of Glycosaminoglycan-Derived Disaccharides

Capillary electrophoresis is a common technique used for glycosaminoglycan-derived disaccharide analysis because of its high resolving power, high separation efficiency, high sensitivity, short analysis time, and straightforward operation. CE coupled to laser-induc ...

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Separation of Chitooligosaccharides in Acidic Solution by Capillary Electrophoresis

Capillary electrophoresis (CE) of chitooligosaccharides (COS) in aqueous solution is effective for their separation from other saccharides. However, COS easily adsorb on negatively charged surfaces, such as fused silica capillaries in acidic solutions. Conventional photo ...

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High-Throughput Capillary ElectrophoresisMass Spectrometry: From Analysis of Amino Acids to Analysis of Protein Complexes

Recent advances in capillary electrophoresis–mass spectrometry (CE-MS) interfacing using porous tip is leading to commercialization of CE-MS with a sheathless interface for the first time. The new sheathless interface in conjunction with CE capillary coatings using self-coat ...

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Fluorescence-Based Technique for Analyzing Retinoic Acid

Retinoic acid (RA) is a potent transcriptional activator whose actions are mediated by members of the nuclear hormone receptor family. In addition to playing key roles in embryonic development and in tissue maintenance in the adult, RA is a potent anticarcinogenic agent currently in clini ...

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Reverse-Phase High-Performance Liquid Chromatography (HPLC) Analysis of Retinol and Retinyl Esters in Mouse Serum and Tissues

The ability to measure endogenous metabolites of retinoids (vitamin A and its derivatives) in biological samples is key to understanding the crucial physiological actions of vitamin A. Over the years, many assays and methods have been developed to analyze different retinoids in biolog ...

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Visualization of Retinoid Storage and Trafficking by Two-Photon Microscopy

Vertebrate vision is maintained by the retinoid (visual) cycle, a complex enzymatic pathway that operates in the retina to regenerate the visual chromophore, 11-cis-retinal, a prosthetic group of rhodopsin that undergoes activation by light. Many different mutations in genes encod ...

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Molecular Biology and Analytical Chemistry Methods Used to Probe the Retinoid Cycle

The retinoid (visual) cycle is a complex enzymatic pathway essential for regeneration of the visual chromophore, 11-cis-retinal, a component of rhodopsin that undergoes activation by light in vertebrate eyes. Pathogenic mutations within genes encoding proteins involved in the re ...

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Assay of Retinol-Binding ProteinTransthyretin Interaction and Techniques to Identify Competing Ligands

The principles of fluorescence resonance energy transfer have been utilized to develop a high-throughput assay which detects compounds that interfere with interaction between retinol-binding protein (RBP) and transthyretin (TTR). In this assay, the intrinsic fluorescence f ...

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The Interaction Between Retinol-Binding Protein and Transthyretin Analyzed by Fluorescence Anisotropy

The retinol carrier retinol-binding protein (RBP) forms in blood a complex with the thyroid hormone carrier transthyretin (TTR). The interactions of retinoid–RBP complexes, as well as of unliganded RBP, with TTR can be investigated by means of fluorescence anisotropy. RBP represents the ...

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Detection of Retinoic Acid Catabolism with Reporter Systems and by In Situ Hybridization for CYP26 Enzymes

Retinoic acid (RA), an active form of vitamin A, is essential for life in vertebrates, owing to its capacity of influencing expression of a sizable fraction of all genes and proteins. It functions via two modes: (1) as controlling ligand for specific transcription factors in the nucleus it stimulat ...

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Diet in Vitamin A Research

A properly formulated diet is an essential underpinning for all in vivo research. This chapter focuses on the use of diet in retinoid research from two perspectives: human research, in which diet is usually variable and analysis of dietary intake is paramount to interpreting the study’s result ...

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Analysis of the Retinoid Isomerase Activities in the Retinal Pigment Epithelium and Retina

Light sensitivity in the vertebrate retina is mediated by the opsin visual pigments inside rod and cone photoreceptor cells. These pigments consist of a G protein-coupled receptor and the photo-sensitive ligand, 11-cis-retinaldehyde (11-cis-RAL). Absorption of a photon by an opsin pig ...

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