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细胞功能测定

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Peptidomics: Divide et Impera

The term “peptidomics” can be defined as the systematic analysis of the peptide content within a cell, organelle, tissue or organism. The science of peptidomics usually refers to the studies of naturally occurring peptides. Another meaning refers to the peptidomics approach to protein an ...

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Mass Spectrometric Analysis of Molluscan Neuropeptides

The central nervous systems of molluscan species contain high levels of structurally diverse peptides that function as neurotransmitters, neuromodulators or neurohormones. Peptide diversity is believed to be a way to increase the information handling capacity of neurons in the c ...

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Monitoring Neuropeptides In Vivo via Microdialysis and Mass Spectrometry

Neuropeptides are important signaling molecules that regulate many essential physiological processes. Microdialysis offers a way to sample neuropeptides in vivo. When combined with liquid chromatography–mass spectrometry detection, many known and unknown neuropept ...

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Protocols for Peptidomic Analysis of Spider Venoms

Spider venom contains a complex mixture of components with a large range of molecular masses (0.1–60kDa) exhibiting a diverse array of actions. Most of these components are proteinaceous molecules – biologically active proteins and peptides. Proteomics profiling of spider venoms (t ...

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Purification and Characterization of Biologically ctive Peptides from Spider Venoms

Spider venoms represent invaluable sources of biologically active compounds suitable for use in life science research and also having a significant potential for biotechnology and therapeutic applications. The methods reported herewith are based on our long experience of spid ...

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MALDI-TOF Mass Spectrometry Approaches to the Characterisation of Insect Neuropeptides

The diversity of insect neuropeptides coupled with the limitations from the small size of the insects themselves combine to make positive identification through peptide sequencing a highly challenging task. The advent of the “soft-ionisation” techniques of MALDI-TOF and electro ...

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Affinity Peptidomics: Peptide Selection and Affinity Capture on Hydrogels and Microarrays

Affinity peptidomics relies on the successfully proven approach used widely in mass-spectrometry-based protein analysis, where protein samples are proteolytically digested prior to the analysis. Unlike traditional proteomic analyses, affinity peptidomics employs a ...

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Immunochemical Methods for the Peptidomic Analysis of Tachykinin Peptides and Their Precursors

The tachykinins represent the largest known peptide family and are responsible for a range of pleiotropic functions in both vertebrates and invertebrates. Recent research has shown a diversity of mechanisms such as mRNA splicing, precursor processing and post-translation modif ...

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Peptidomic Approaches to the Identification and Characterization of Functional Peptides in Hydra

Little is known about peptides that control developmental processes such as cell differentiation and pattern formation in metazoans. The cnidarian Hydra is one of the most basal metazoans and is a key model system for studying the peptides involved in these processes. We developed a novel pep ...

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Bioinformatic Identification of Plant Peptides

Plant peptides play a number of important roles in defence, development and many other aspects of plant physiology. Identifying additional peptide sequences provides the starting point to investigate their function using molecular, genetic or biochemical techniques. Due to their ...

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Analysis of Phospholipids by High Performance Liquid Chromatography

Phospholipids are amphipathic molecules of great importance and widespread in biological material. They play an essential role in the structure and function of biological membranes. They are classified according to the nature of the “backbone” residue (glycerol or sphingosine), t ...

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Isolation of Cholinergic-Specific Synaptosomes by Immunoadsorption

The most widely used subcellular fractionation techniques (e.g., centrifugation) are dependent on differences in physical parameters between organelles (e.g., size and density). In contrast, immunological techniques rely on biological differences, i.e., the expression of dif ...

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Isolation and Purification of Functionally Intact Mitochondria from Plant Cells

The isolation of mitochondria from plant cells that display similar biochemical and morphological characteristics to those observed in vivo requires considerable expertise. Although it is relatively easy to prepare a crude mitochondrial fraction by differential centrifu ...

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Separation and Analysis of Phospholipids by Thin Layer Chromatography

The membranes of animal cells contain a substantial amount of lipid, which may account for between 20 and 80% of the membrane mass. The predominant lipid components are phospholipids (PL) and cholesterol; with glycolipids, cholesterol esters, glycerides, and free fatty acids as minor const ...

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Measurement of Cholesterol in Membranes

Cholesterol is an amphipathic molecule of great importance in biology. It can be measured by enzymic and chemical methods, directly or indirectly following extraction from tissues or membranes. Before determination, cholesterol esters must be hydrolyzed using either chemical me ...

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Chemical Assays for Proteins

The choice of a protein assay for membranes and membrane proteins is dependent on a number of considerations: sensitivity, specificity (both with respect to variation between proteins and interference by nonprotein components), and simplicity.

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Analysis of Fatty Acids by Gas Liquid Chromatography

The fatty acids (FA) of animal, plant, and microbial origin are predominantly unbranched aliphatic chains with an even number of carbon atoms and a single carboxyl group. FA can be classified as saturated, monounsaturated, or polyunsaturated (Fig. 1). Saturates have all single carbon-to-c ...

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Separation and Analysis of Membrane Proteins by SDS-Polyacrylamide Gel Electrophoresis

Polyacrylamide gel electrophoresis (PAGE) in the presence of the anionic detergent, sodium dodecyl sulfate (SDS), is probably the most commonly used technique for the analysis of protein mixtures. SDS is a very effective solubilizing agent for a wide range of polypeptides, including mem ...

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The Identification of Subcellular Fractions from Mammalian Cells

The chapters in the two volumes of Biomembrane Protocols are broadly devoted to the methodologies involved in the structural and functional characterization of cell membranes. In many cases, a study of a particular membrane presupposes that it can be isolated by disruption of the cells and fr ...

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Quantification of Human Cytomegalovirus by Competitive PCR and Capillary Electrophoresis

Human cytomegalovirus (CMV) is a ubiquitous pathogen found in 40–100% of adults, and in about 1% of live births in the United States (1). It is the most common fetal and perinatal infectious organism; approx 10% of infected neonates are born with symptomatic congenital CMV disease, which is the most comm ...

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