• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biochemistry

Cloud-point PEG Glass Surfaces for Imaging of Immobilized Single Molecules by Total-internal-reflection Microscopy Authors:  Zhengjian Zhang, Sang Ryul Park, Alexandros Pertsinidis and Andrey Revyakin, date: 04/05/2016, view: 6399, Q&A: 0
This effective, robust protocol generates glass coverslips coated with biotin-functionalized polyethylene glycol (PEG), making the glass surface resistant to non-specific absorption of biomolecules, and permitting immobilization of biomolecules for subsequent single-molecule tracking of biochemical reactions. The protocol can be completed in one ...
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Cancer Biology

Colon Cancer-associated Fibroblast Establishment and Culture Growth Authors:  Mercedes Herrera, Alberto Herrera, María Jesús Larriba, Gemma Ferrer-Mayorga, Antonio García de Herreros, Félix Bonilla, Josep Baulida and Cristina Peña, date: 04/05/2016, view: 6769, Q&A: 0
Cancer-associated fibroblasts (CAFs) are one of the major players in tumor-stroma crosstalk. Findings in experimental studies suggest important roles for CAFs in regulation of tumor growth, metastasis and drug response (Hanahan and Coussens, 2012). Furthermore, their clinical relevance is supported by new findings from tumor analyses, ...
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Extraction and Quantification of Tryptophan and Kynurenine from Cultured Cells and Media Using a High Performance Liquid Chromatography (HPLC) System Equipped with an Ultra-sensitive Diode Array Detector Authors:  Jeffrey Kim, Benjamin Stewart and Robert H Weiss, date: 04/05/2016, view: 5082, Q&A: 0
Evidence of the involvement of tryptophan and its metabolite, kynurenine, in various biological processes including cancer is constantly expanding. Analysis of cell extracts and culture media can allow for quick snapshots of the metabolic fluctuations occurring in vitro. Here, we describe a method for metabolite extraction from mammalian ...
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Microbiology

Rust Removal Experiments Authors:  Mahmoud Kamal Ahmadi and Blaine A. Pfeifer, date: 04/05/2016, view: 10213, Q&A: 0
Iron oxidation, known as rust formation, causes enormous loss in term of property damages and associated economic risks. Depending on the degree of formation, rust consists of several layers of iron in different oxidation states. The brownish top layer is mostly iron (III) oxide-hydroxide [FeO(OH), Fe(OH)3] while the deepest black ...
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Methods for Detecting Microbial Methane Production and Consumption by Gas Chromatography Authors:  Jared T. Aldridge, Jennie L. Catlett, Megan L. Smith and Nicole R. Buan, date: 04/05/2016, view: 5758, Q&A: 0
Methane is an energy-dense fuel but is also a greenhouse gas 25 times more detrimental to the environment than CO2. Methane can be produced abiotically by serpentinization, chemically by Sabatier or Fisher-Tropsh chemistry, or biotically by microbes (Berndt et al., 1996; Horita and Berndt, 1999; Dry, 2002; Wolfe, 1982; Thauer, ...
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Procedure for Rhamnolipids Quantification Using Methylene-blue Authors:  Tsiry Rasamiravaka, Olivier M. Vandeputte and Mondher El Jaziri, date: 04/05/2016, view: 8252, Q&A: 2
Rhamnolipids produced by Pseudomonas aeruginosa (P. aeruginosa) represent a group of biosurfactants with various applications (e.g., bioremediation of oil spills, cosmetics, detergents and cleaners). The commonly used colorimetric methods for rhamnolipid quantification, including anthrone, phenol−sulfuric acid and ...
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Molecular Biology

Conjugation of Duplexed siRNN Oligonucleotides with DD-HyNic Peptides for Cellular Delivery of RNAi Triggers Authors:  Alexander S. Hamil, Khirud Gogoi and Steven F. Dowdy, date: 04/05/2016, view: 4952, Q&A: 0
Despite the great promise that short interfering RNA (siRNA) induced RNAi responses hold as a therapeutic modality, due to their size (~15 kDa) and high negative charge (Bumcrot et al., 2006), siRNAs have no bioavailability and require a delivery agent to enter cells (Figure 1). TAT peptide transduction domain (PTD) has been developed as ...
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Neuroscience

Measurement of Inositol Triphosphate Levels from Rat Hippocampal Slices Authors:  Nino Tabatadze and Catherine Woolley, date: 04/05/2016, view: 4948, Q&A: 0
Inositol triphosphate (IP3) is an important second messenger that participates in signal transduction pathways in diverse cell types including hippocampal neurons. Stimulation of phospholipase C in response to various stimuli (hormones, growth factors, neurotransmitters, neurotrophins, neuromodulators, odorants, light, etc.) ...
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Isolation and Primary Cell Culture of Mouse Dorsal Root Ganglion Neurons Authors:  Theresa Heinrich, Christian A. Hübner and Ingo Kurth, date: 04/05/2016, view: 15755, Q&A: 0
We here provide a detailed protocol for the isolation and culture of primary mouse sensory neurons. The cell bodies of sensory afferent pseudounipolar neurons are located in dorsal root ganglia (DRGs) along the vertebral column. Dissected mouse DRGs can be dissociated into single cells by enzymatic digestion to obtain primary cultures of mouse ...
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Plant Science

Preparation of Mitotic and Meiotic Metaphase Chromosomes from Young Leaves and Flower Buds of Coccinia grandis Authors:  Sangram Sinha, Kanika Karmakar, Ravi S. Devani, Jayeeta Banerjee, Rabindra K. Sinha and Anjan K. Banerjee, date: 04/05/2016, view: 7072, Q&A: 1
Somatic chromosomes are usually studied from the root tip cells of the plants for cytological investigations. In dioecious plant species like Coccinia grandis, it is very difficult to get meristematic root tip cells from the mature plants of the respective sex forms. In this report, young leaves of the respective sexual phenotypes were ...
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Strategies for Performing Dynamic Gene Perturbation Experiments in Flowers Authors:  Diarmuid S. Ó’Maoiléidigh, Emmanuelle Graciet and Frank Wellmer, date: 04/05/2016, view: 4680, Q&A: 0
Dissecting the gene regulatory networks (GRNs) underlying developmental processes is a central goal in biology. The characterization of the GRNs underlying flower development has received considerable attention, however, novel approaches are required to reveal temporal and spatial aspects of these GRNs. Here, we provide an overview of the options ...
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Analysis of Telomeric G-overhangs by in-Gel Hybridization Authors:  Sona Valuchova, Elisa Derboven and Karel Riha, date: 04/05/2016, view: 4304, Q&A: 0
Telomeric DNA in majority of eukaryotes consists of an array of TG-rich tandem repeats. The TG-rich DNA strand is oriented with its 3’ end towards chromosome termini and is usually longer than its complementary CA-rich strand, thus forming 3’ single stranded overhang (G-overhang). G-overhangs arise from incomplete replication of chromosome termini ...
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Total RNA Extraction from Grape Berry Skin for Quantitative Reverse Transcription PCR and Microarray Analysis Authors:  Mami Suzuki and Katsuhiro Shiratake, date: 04/05/2016, view: 4740, Q&A: 0
Extraction of high quality RNA is an essential step for quantitative reverse transcription PCR (qRT-PCR) and microarray analysis. However, it is not easy to extract high quality RNA from fruit materials, which contain high amounts of polysaccharides, lipids and secondary metabolites. Wan and Wilkins (1994) had developed ‘Hot Borate Method’ to ...
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Expression, Purification and Enzymatic Assay of Plant Histone Deacetylases Authors:  Ming Luo and Keqiang Wu, date: 04/05/2016, view: 4980, Q&A: 0
Histone deacetylases (HDACs) catalyzing the removal of acetyl groups from lysine residues of histone and non-histone proteins play vital roles in regulation of gene transcription. In plants, HDACs can be grouped into three families, including RPD3-type, SIR2-type and plant specific HD2-type HDACs. Here we describe a method to determine plant HDAC ...
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Stem Cell

Isolation and Culture of Human CD133+ Non-adherent Endothelial Forming Cells Authors:  Michaelia P. Cockshell and Claudine S. Bonder, date: 04/05/2016, view: 6584, Q&A: 0
Circulating endothelial progenitor cells (EPCs) have been the focus of many clinical trials due to their roles in revascularisation following ischemic events such as acute myocardial infarction as well as their contribution to vascular repair during organ transplantation. Research on EPCs has been controversial due to the lack of distinct markers ...
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