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Cell Biology

Flow Cytometric Detection of Mitochondrial Membrane Potential Authors:  Hsin-Yi Chang, Hsuan-Cheng Huang, Tsui-Chin Huang, Pan-Chyr Yang, Yi-Ching Wang and Hsueh-Fen Juan, date: 04/20/2013, view: 12734, Q&A: 1
Mitochondrial membrane potential (Δψm) is an important parameter of mitochondrial function and an indicator of cell health. Depletion of Δψm suggests the loss of mitochondrial membrane integrity reflecting the initiation of the proapoptotic signal. Recently, lipophilic cationic fluorescent dyes have been developed to detect Δψm by accumulating in ...
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Flow Cytometric Detection of Reactive Oxygen Species Authors:  Hsin-Yi Chang, Hsuan-Cheng Huang, Tsui-Chin Huang, Pan-Chyr Yang, Yi-Ching Wang and Hsueh-Fen Juan, date: 04/20/2013, view: 19339, Q&A: 1
Reactive oxygen species (ROS) are molecules containing hydroxyl radicals or peroxides with unpaired electrons. In healthy aerobic cells, ROS are produced naturally as a byproduct of oxidative phosphorylation, oxidoreductase enzymes, or metal catalyzed oxidation at a controlled rate. However, ROS can be induced under some stress conditions ...
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Immunology

Evaluation of Caspase-1 Activation and IL-1β Production in A Kainic Acid Microdyalisis Brain Injury Model Authors:  Antonio S. Herranz, Eulalia Bazán, Diana Reimers, María T. Montero-Vega, Adriano Jménez-Escrig and Pablo Pelegrín, date: 04/20/2013, view: 4611, Q&A: 0
Intracerebral infusion of kainic acid (KA) by a microdialysis probe induces a focal swelling in the brain-perfused area which promotes inflammation (Compan et al., 2012; Oprica et al., 2003). The microdialysis technique allows the local in vivo perfusion of KA and the simultaneous collection of inflammatory mediators, ...
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Microbiology

Plasmodium falciparum Rosette Disruption Assay Authors:  Micheline Guillotte, Odile Mercereau-Puijalon and Inès Vigan-Womas, date: 04/20/2013, view: 5127, Q&A: 0
Rosetting, i.e. the capacity of Plasmodium falciparum-infected red blood cells (iRBCs) to bind two or more uninfected red blood cells (RBCs) is associated with severe malaria in African children. Disruption of rosettes using small soluble inhibitors or specific antibodies is viewed as an interesting strategy to treat or prevent ...
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Plasmodium falciparum Rosette Formation Assay Authors:  Inès Vigan-Womas, Micheline Guillotte and Odile Mercereau-Puijalon, date: 04/20/2013, view: 7007, Q&A: 0
Rosetting, i.e. the capacity of red blood cells (iRBCs) infected with mature parasite stages to bind two or more uninfected red blood cells (RBCs) is a virulence factor of Plasmodium falciparum. This protocol describes an in vitro assay to monitor rosette formation by P. falciparum-infected red blood cells, ...
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Packaging of Retroviral RNA into Viral Particles Analyzed by Quantitative Reverse Transcriptase-PCR Authors:  Bianca Hoffmann and Bastian Grewe, date: 04/20/2013, view: 7847, Q&A: 0
Formation of viral particles and packaging of genomic retroviral RNA into these particles are important steps in the late phase of the viral replication cycle. The efficiency of the incorporation of viral or cellular RNAs into viral particles can be studied using a quantitative Reverse Transcriptase-PCR (RT-qPCR)-based approach. After isolation of ...
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Molecular Biology

High-throughput Method for Determination of Seed Paternity by Microsatellite Markers Authors:  Samik Bhattacharya and Ian T. Baldwin, date: 04/20/2013, view: 6309, Q&A: 0
In this protocol, determination of seed paternity by microsatellite markers in Nicotiana attenuata is described. However, this does not include a protocol for the novel marker selection/identification, but rather exploits the markers generated for a closely related species N. tabacum (Bindler et al., 2007). This is a ...
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Detection and Cloning of Spliced Transcripts by RT-PCR Authors:  Bianca Hoffmann and Bastian Grewe, date: 04/20/2013, view: 6168, Q&A: 0
Using a Reverse Transcriptase-PCR approach spliced transcripts can be converted to cDNA, amplified and cloned into an expression plasmid. Sequencing of the obtained cDNA allows identification of the splicing events that generated the detected RNA (Grewe et al., 2012).
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p65 Chromatin Immunoprecipitation Protocol Author:  Crissy Dudgeon, date: 04/20/2013, view: 4950, Q&A: 0
Chromatin Immunoprecipitation (ChIP) is an important procedure that allows you to verify if a certain protein is physically located at a regulatory region. This information, taken together with other procedures such as luciferase assays and EMSAs, will give definitive proof that the query protein is involved in the transcription of a protein. ...
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Neuroscience

Primary Culture of Cortical Neurons Authors:  Rieko Muramatsu and Toshihide Yamashita, date: 04/20/2013, view: 9741, Q&A: 1
Primary culture of neurons from cerebral cortex is a popular model to study neuronal function in vitro and to explore the molecular mechanism of neurite outgrowth in the developing and adult central nervous system. This protocol is for preparing a culture of cerebral cortical neurons from postnatal rodent brain (Muramatsu et al., ...
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Culture of Rat Olfactory Ensheathing Cells Using EasySep® Magnetic Nanoparticle Separation Authors:  Susan Louise Lindsay and Susan Carol Barnett, date: 04/20/2013, view: 4256, Q&A: 0
Olfactory ensheathing cells (OECs) can be isolated and purified from a range of postnatal day 7-day to 10-day rat olfactory bulbs. Rat OECs express the CD271/p75NTR receptor and using the “Do-It-Yourself” magnetic nanoparticle EasySep kit from STEMCELL technologies this protocol allows the selective purification of these cells in less than 50 min. ...
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Plant Science

Determination of Nectar Nicotine Concentration in N. attenuata Authors:  Eva Rothe, Matthias Schöttner, Danny Kessler and Ian T. Baldwin, date: 04/20/2013, view: 4957, Q&A: 0
In this protocol, the determination of the nicotine concentration in nectar of Nicotiana attenuata is described. This method is applicable for the investigation of small amounts of nectar (above 1 μl). It is a high-throughput protocol optimized and streamlined for one skilled person to process approximately 100 nectar samples per day.
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