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Cancer Biology

Detection of ALT Associated Promyelocytic Leukemia Nuclear Bodies (APBs) by Immunofluorescence-FISH (IF-FISH) Authors:  Siamak A. Kamranvar and Maria G. Masucci, date: 12/05/2014, view: 6553, Q&A: 0
The activation of functions that counteract the physiological shortening of telomeres in rapidly proliferating cell is prerequisite for the progression of cancer cells to full malignancy (Collado et al., 2007). In most human cancers, the length of telomere is maintained through up-regulation of telomerase whereas a telomerase-independent ...
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Cell Biology

TGFβ Stimulation Assay Author:  Emanuel Rognoni, date: 12/05/2014, view: 6122, Q&A: 0
TGFβ is part of a growth factor superfamily which modulates cell growth, differentiation, adhesion, migration, ECM synthesis and apoptosis (Massague, 1998; Siegel and Massague, 2003). Free TGFβ binds to its high affinity TGFβ receptor, a receptor serine/threonine kinase, inducing phosphorylation of Smad2/3 which subsequently forms a complex with ...
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TGFβ Release Co-culture Assay Author:  Emanuel Rognoni, date: 12/05/2014, view: 4630, Q&A: 0
TGFβ is a potent cytokine modulating various processes including proliferation, differentiation, ECM synthesis and apoptosis (Siegel and Massague, 2003). Thus in many tissues availability of TGFβ is tightly regulated. TGFβ is secreted as an inactive complex where it is encapsulated by the latency associated protein (LAP), a ligand trap protein, ...
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Microbiology

Visualization of Cell Complexity in the Filamentous Cyanobacterium Mastigocladus laminosus by Transmission Electron Microscopy (TEM) Authors:  Dennis J. Nürnberg, Giulia Mastroianni, Conrad W. Mullineaux and Graham D. McPhail, date: 12/05/2014, view: 4586, Q&A: 0
The cyanobacterium Mastigocladus laminosus (M. laminosus) is one of the most morphologically complex prokaryotes. It forms long chains of cells that are connected via septal junction complexes; such complexes allow diffusion of metabolites and regulators between neighboring cells. Cellular division occurs in multiple planes, ...
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Induction of Tigecycline Resistance in Acinetobacter baumannii Authors:  Ming-Feng Lin, Yun-You Lin and Chung-Yu Lan, date: 12/05/2014, view: 3999, Q&A: 0
Multidrug resistance Acinetobacter baumannii (A. baumannii) (MDRAB) has emerged as a serious threat in hospitals in recent years. Currently, there are few antibiotics, including tigecycline, available to treat infections caused by MDRAB effectively. Both tigecycline-resistant and tigecycline-susceptible isogenic strains of MDRAB ...
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Minimal Inhibitory Concentration (MIC) Assay for Acinetobacter baumannii Authors:  Ming-Feng Lin, Yun-You Lin and Chung-Yu Lan, date: 12/05/2014, view: 15973, Q&A: 2
Minimal inhibition concentration (MIC) is the lowest concentration of an antimicrobial agent that can inhibit the visible growth of a microorganism after overnight incubation. MIC determination is used as not only a diagnostic tool in treating bacterial infections for clinicians but also a research method in evaluating the efficacy of an ...
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Enzymatic Activity Assays in Yeast Cell Extracts Authors:  Melike Çağlayan and Samuel H. Wilson, date: 12/05/2014, view: 5783, Q&A: 0
Saccharomyces cerevisiae (S. cerevisiae) (commonly known as baker’s yeast) is a model organism that has a similar upstream base excision repair (BER) pathway for the repair of methylated bases as that in mammalian cells, and it is very easy to maintain in the laboratory environment. Here, we described a method to prepare cell ...
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Neuroscience

Fluorescent Measurement of Synaptic Activity Using SynaptopHluorin in Isolated Hippocampal Neurons Authors:  Hongmei Li, Han-A Park and Elizabeth A. Jonas, date: 12/05/2014, view: 5406, Q&A: 0
This protocol comprises the entire process of fluorescent measurement of vesicle recycling using the probe SynaptopHluorin, a pH-dependent GFP variant whose fluorescence increases at the synapse upon vesicle release due to fluorescence quenching in acidic vesicles. This technique provides a genetic tool to monitor synaptic vesicle recycling in ...
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Creating a Rat Model of Chronic Variate Stress Authors:  Rocío M. de Pablos, Manuel Sarmiento and Ana María Espinosa-Oliva, date: 12/05/2014, view: 5099, Q&A: 0
Stress is a condition of human experience and an important factor in the onset of various diseases. There are numerous studies showing how stress can accelerate cell aging, immune senescence and some age-related diseases such as neurodegenerative disorders and osteoporosis. However, the effects of stress have different consequences depending on ...
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Plant Science

In situ Digestion of Wheat Cell-wall Polysaccharides Authors:  Dušan Veličković and Hélène Rogniaux, date: 12/05/2014, view: 4857, Q&A: 2
Cell walls of the wheat endosperm are mostly composed of arabinoxylans (AX) and mixed (1→3), (1→4)-β-glucans (BG) (Saulnier et al., 2012). Here, we present an optimized protocol to degrade enzymatically these cell-wall polysaccharides into oligosaccharides, directly from wheat grain cross sections. The main difficulty is to provide a ...
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Vanillin Assay of Arabidopsis Seeds for Proanthocyanidins Authors:  Lijie Xuan, Zhong Wang and Lixi Jiang, date: 12/05/2014, view: 4160, Q&A: 0
Proanthocyanidins (PAs) are colorless flavonoid polymers and deposit in Arabidopsis seed coat specifically. Oxidation of PAs gives rise to brown color of mature seeds. PA accumulation can be affected by a number of growth conditions, such as temperature and sun light. PAs, which are converted from anthocyanidins, can protect seeds from ...
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CytoTrap Two-Hybrid Screening Assay Authors:  Lei Zhang, Liqun Du and B. W. Poovaiah, date: 12/05/2014, view: 5774, Q&A: 0
CytoTrap two-hybrid system provides an alternate strategy to detect protein-protein interactions in yeast. In this system, bait protein is fused with human son of sevenless (hSos) protein (Li et al., 1993), and a cDNA library or prey protein is expressed by fusion with myristoylation signal which anchors the prey fusion protein to yeast ...
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Infection Assays of Tomato and Apple Fruit by the Fungal Pathogen Botrytis cinerea Authors:  Zhanquan Zhang, Guozheng Qin, Boqiang Li and Shiping Tian, date: 12/05/2014, view: 5466, Q&A: 0
Botrytis cinerea (B. cinerea) is an aggressive fungal pathogen that infects more than 200 plant species. Furthermore, the pathogen can attack fruits of some plants, such as tomato and apple. B. cinerea has become one of the model systems in molecular phytopathology because of its economic importance and sophisticated ...
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Purification of Starch Granules from Arabidopsis Leaves and Determination of Granule-Bound Starch Synthase Activity Authors:  Tomás Albi, M. Isabel Ortiz-Marchena, M. Teresa Ruiz, José M. Romero and Federico Valverde, date: 12/05/2014, view: 4338, Q&A: 0
Starch constitutes the most important carbon reserve in plants and is composed of branched amylopectin and linear amylose. The latter is synthesized exclusively by the Granule-Bound Starch Synthase (GBSS, EC 2.4.1.21). Here we report a readily reproducible, specific and highly sensitive protocol, which includes the isolation of intact starch ...
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Determination of Soluble Sugars in Arabidopsis thaliana Leaves by Anion Exchange Chromatography Authors:  M. Isabel Ortiz-Marchena, M. Teresa Ruiz, Federico Valverde and José M. Romero, date: 12/05/2014, view: 6478, Q&A: 0
Determination of soluble sugars is basic for the study of carbon metabolism in plants. Soluble sugar quantitation can be achieved by enzymatic methods implying different coupled reactions. Here we describe a simple method that allows rapid determination of the most abundant soluble sugars (glucose, fructose and sucrose) in Arabidopsis ...
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