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细胞功能测定

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Imaging of Intracellular Hydrogen Peroxide Production with Hyper Upon Stimulation of Hela Cells with Egf

Reactive oxygen species (ROS) regulate both normal cell functions by activating a number of enzymatic cascades and pathological processes in many diseases by inducing oxidative stress. For many years since the discovery of ROS in biological systems, there were no adequate methods of det ...

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Measuring Redox Changes In Vivo in Leaves: Prospects and Technical Challenges

In leaves, the functioning of many key proteins under conditions promoting oxidative stress depends to a large extent on the redox potential of the glutathione couple. Routine measurements of the glutathione pool in leaves are destructive and labor-intensive processes that tend to und ...

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Redox-Sensitive Green Fluorescent Protein: Probes for Dynamic Intracellular Redox Responses. A Review

The quantification of transient redox events within subcellular compartments, such as those involved in certain signal transduction pathways, requires specific probes with high spatial and temporal resolution. Redox-sensitive variants of the green fluorescent protein (r ...

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Detection and Measurement of Reactive Oxygen Intermediates in Mitochondria and Cells

Reactive oxygen intermediates (ROIs) play a key role in a number of human diseases either by inducing cell death, cellular proliferation, or by acting as mediators in cellular signaling. Therefore, their measurement in vivo and in cell culture is desirable but technically difficult and oft ...

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The Measurement of Nitric Oxide and Its Metabolites in Biological Samples by Ozone-Based Chemiluminescence

A plethora of publications on techniques and methodologies for measuring nitric oxide (NO) or reaction products of NO (NO metabolites) has served in recent years to complicate and confuse the majority of researchers interested in this field. Here, we provide a practical approach and summar ...

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The Role of Redox in Signal Transduction

Functioning and efficient cell signaling is vital for the survival of cells. Over the course of many years, various components have been identified and recognized as crucial for the transduction of signals in cells. Many of the mechanisms allow for a relatively rapid switching of signals, on or of ...

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Generation and Detection of S-Nitrosothiols

Nitric oxide (NO) plays a pivotal role in cellular signaling in many different organisms as the result of the modification of protein activities/functions by protein S-nitrosylation. This NO-dependent posttranslational modification is based on the attachment of NO to the sulfur moi ...

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Analysis of Redox Relationships in the Plant Cell Cycle: Determinations of Ascorbate, Glutathione and Poly (ADPribose) Polymerase (PARP) in Plant Cell

Reactive oxygen species (ROS) and low molecular weight antioxidants, such as glutathione and ascorbate, are powerful signaling molecules that participate in the control of plant growth and development, and modulate progression through the mitotic cell cycle. Enhanced reactive o ...

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Protein-Thiol Oxidation, From Single Proteins to Proteome-Wide Analyses

Protein-thiol oxidation subserves multiple biological functions, from enzymatic catalysis to protein oxidative folding, protein trafficking, reactive oxygen (ROS) and nitrogen (RNS) species sensing and signaling and, more generally, protein redox regulation. Protein- ...

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Analysis of Global and Specific Changes in the Disulfide Proteome Using Redox 2D-PAGE

Protein cysteine sulfhydryl groups are susceptible to a number of redox-dependent modifications, including an interchange between the reduced sulfhydryl and an oxidized disulfide state. A growing body of evidence suggests that reversible disulfide bond formation alters the s ...

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Detection of Carbonylated Proteins in 2-D SDS Page Separations

Protein carbonyls are an index of protein oxidation which, in turn, reflects the interplay of oxidative stress and degradation of oxidatively modified proteins. Protein carbonyls are increased in brain proteins in aging and age-related neurodegenerative disorders, including A ...

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Redox Regulation and Trapping Sulphenic Acid in the Peroxide Sensitive Human Mitochondrial Branched Chain Aminotransferase

The human branched chain aminotransferase enzymes are key regulators of glutamate metabolism in the brain and are among a growing number of redox-sensitive proteins. Studies that use thiol-specific reagents and electrospray ionization mass spectrometry demonstrate that the m ...

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Differentiation of Dendritic Cells from Human Embryonic Stem Cells

Improving our understanding of the interactions between human dendritic cells (DCs) and T cells may contribute to the development of therapeutic strategies for a variety of immune-mediated disorders. The possibility of using DCs themselves as tools to manipulate immune responses o ...

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In Vivo Evaluation of Putative Hematopoietic Stem Cells Derived from Human Pluripotent Stem Cells

Efficient derivation and isolation of hematopoietic stem cells (HSCs) from human pluripotent stem cell (hPSC) populations remains a major goal in the field of developmental hematopoiesis. These enticing pluripotent stem cells (comprising both human embryonic stem cells and ind ...

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Methods for the Derivation and Use of Cardiomyocytes from Human Pluripotent Stem Cells

The availability of human cardiomyocytes derived from embryonic stem cells (ESCs) has generated �considerable excitement, as these cells are an excellent model system for studying myocardial development and may have eventual application in cell-based cardiac repair. Cardiom ...

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Directed Differentiation of Dopamine Neurons from Human Pluripotent Stem Cells

Midbrain dopaminergic (mDA) neurons play a critical role in regulating postural reflexes and movement as well as modulating psychological processes. Dysfunction or degeneration of mDA neurons is involved in a number of neurological disorders including Parkinson’s disease. Ava ...

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Derivation of Oligodendrocyte Progenitor Cells from Human Embryonic Stem Cells

The directed differentiation of human pluripotent stem cells into specific, determined, and high-purity cell types can provide a means to study the cellular and molecular mechanisms of development and to generate cells for potential therapeutic applications. The ability to derive h ...

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The Generation of Embryoid Bodies from Feeder-Based or Feeder-Free Human Pluripotent Stem Cell Cultures

Embryoid body (EB) formation is a traditional method of inducing differentiation of pluripotent stem cells (PSCs). It is a routine in vitro test of pluripotency as well as the first stage in many differentiation protocols targeted toward the production of a specific lineage or cellular popu ...

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Episomal Transgene Expression in Pluripotent Stem Cells

Herpes simplex type 1 (HSV-1) amplicon vectors possess a number of features that make them excellent vectors for the delivery of transgenes into stem cells. HSV-1 amplicon vectors are capable of efficiently transducing both dividing and nondividing cells and since the virus is quite large, 1 ...

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Gene Targeting in Human Pluripotent Stem Cells

Targeted homologous recombination (HR) is an essential tool in stem cell biology. It can be used to study gene function and is a highly developed technology in the mouse where precise genetic modifications are introduced into the genome via HR in mouse embryonic stem cells (mESCs). However, gene t ...

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