• 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

Cancer Biology

Bone Resorption Assay Authors:  Carina Scholtysek, Gerhard Krönke and Georg Schett, date: 07/20/2014, view: 9789, Q&A: 0
The Bone resorption assay provides an easy to use protocol for quantitatively measuring in vitro osteoclast-mediated bone resorption. Osteoclasts can be seeded onto the bone slices and formation of resorption pits can be quantified via toluidinblue staining (Scholtysek et al., 2013).
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Developmental Biology

Hedgehog (Hh) Reporter Activity Assay Author:  Chen Zhao, date: 07/20/2014, view: 9767, Q&A: 0
This protocol is for testing responses of a candidate cell line/cell lines to Hh ligands or Hh pathway agonists stimulation. This protocol can also be adapted to screen small molecule libraries or biologics that contain activities to either increase or decrease Hh pathway responses. Canonical Hh signaling activity transcriptionally induces Hh ...
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Wnt Reporter Activity Assay Author:  Chen Zhao, date: 07/20/2014, view: 20550, Q&A: 2
This protocol is for testing responses of a candidate cell line/cell lines to Wnt ligands or Wnt pathway agonists stimulation. This protocol can also be adapted to screen small molecule libraries or biologics that contain activities to either increase or decrease Wnt pathway responses. Canonical Wnt signaling activity transcriptionally induces Wnt ...
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Fluorescence Microscopy for Cilia in Cultured Cells and Zebrafish Embryos Authors:  Jingli Cao, Xueliang Zhu and Xiumin Yan, date: 07/20/2014, view: 10027, Q&A: 0
Cilia are microtubule-based hair-like projections found in organisms, ranging from protozoa to mammals. This protocol provides methods for immunofluorescence staining of cilia in cultured cells and zebrafish embryos.
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Immunology

Isolation of ILC2 from Mouse Liver Authors:  Tamar Mchedlidze and Stefan Wirtz, date: 07/20/2014, view: 7876, Q&A: 0
Group 2 innate lymphoid cells (ILC2) are a recently characterized cell population which lacks specific antigen receptors and contributes to immune responses at mucosal surfaces of lung and gut. Recently, we demonstrated that ILC2 expand in the context of chronic liver diseases in mice and contribute to pathology in an IL-33 dependent manner. Here, ...
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Macrophage Inflammatory Assay Authors:  Thomas J. Bartosh and Joni H. Ylostalo, date: 07/20/2014, view: 20040, Q&A: 0
Macrophages represent a widely distributed and functionally diverse population of innate myeloid cells involved in inflammatory response to pathogens, tissue homeostasis and tissue repair (Murray and Wynn, 2011). Macrophages can be broadly grouped into two subpopulations with opposing activites: M1 or pro-inflammatory macrophages that promote ...
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Microbiology

In vitro EBV Infection of Mononuclear Cells that Have Been Cryo-preserved Authors:  Shanie Saghafian-Hedengren, Ebba Sohlberg, Jakob Theorell, Claudia Carvalho-Queiroz, Noémi Nagy, Jan-Olov Persson, Caroline Nilsson, Yenan T. Bryceson and Eva Sverremark-Ekström, date: 07/20/2014, view: 4730, Q&A: 0
Epstein-Barr Virus (EBV) is a B-lymphotropic herpesvirus which the majority of adult human population is latently-infected with. Various immunological and molecular in vitro studies have been facilitated by the use of EBV’s ability to infect and transform B cells to immortalized polyclonal B cell lines. Many of these studies use freshly ...
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Affinofile Assay for Identifying Macrophage-Tropic HIV-1 Authors:  Sarah B. Joseph, Benhur Lee and Ronald Swanstrom, date: 07/20/2014, view: 5405, Q&A: 0
The ability to enter monocyte-derived macrophage (MDM) in vitro is commonly used to define macrophage-tropic HIV-1 despite the fact that viruses vary continuously in their ability to enter MDMs in vitro, and MDMs vary in their ability to support HIV-1 entry (Joseph et al., 2014; Peters et al., 2006). This makes ...
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Chromatin Fractionation Assay in Fission Yeast Authors:  Tatsuki Kunoh and Toshiyuki Habu, date: 07/20/2014, view: 7041, Q&A: 0
The protein recruitment onto chromatin is a critical process for DNA metabolism, including DNA replication, DNA repair and DNA recombination. Especially DNA modification enzymes and checkpoint proteins are loaded onto DNA damage sites in a context-dependent manner. In our recent study (Kunoh and Habu, 2014), the chromatin association of Pcf1, a ...
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Development of a Novel Assay for Synthesis and Hydrolysis of Sedoheptulose 1,7-bisphosphate (SBP) in vitro by Combinations of Purified Fructose 1,6-bisphosphate aldolases (FBA) Proteins and Fructose 1,6-bisphosphatases (FBPase) Proteins from Bacillus methanolicus MGA3 Authors:  Jessica Stolzenberger, Steffen N. Lindner, Marcus Persicke, Trygve Brautaset and Volker F. Wendisch, date: 07/20/2014, view: 5699, Q&A: 0
Bacillus methanolicus (B. methanolicus) is a Gram-positive, thermotolerant, and facultative methylotrophic bacterium that can use the one-carbon (C1) compound methanol as a source of carbon and energy (Schendel et al., 1990; Arfman et al., 1997; Arfman et al., 1992). B. methanolicus uses the ...
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Dye Release Experiments with Dextran Loaded Vesicles Authors:  Marc-Antoine Sani, Neil M. O’Brien-Simpson and Frances Separovic, date: 07/20/2014, view: 5993, Q&A: 0
Dye release experiments are a widely used method to assess the interactions between membrane-active molecules and lipid membranes. Of particular interest is the ability to assess the degree of the lipid bilayer perturbation by simultaneously encapsulating dye of different sizes, such as dextrans grafted with a chromophore. In this assay, dextran ...
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Bacterial Fluorescent-dextran Diffusion Assay Authors:  Neil M. O’Brien-Simpson, Namfon Pantarat, Katrina A. Walsh, Eric C. Reynolds, Marc-Antoine Sani and Frances Separovic, date: 07/20/2014, view: 5626, Q&A: 0
Antimicrobial peptides are known to disrupt bacterial membranes allowing solutes to flow across the membrane in an unregulated manner resulting in death of the organism. Disrupting the bacterial membrane would thus perturb the cells osmotic balance resulting in an initial influx of the external aqueous buffer. We have designed an assay to ...
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Plant Science

Rice Meiotic Chromosome Spread Preparation of Pollen Mother Cells Authors:  Xingwang Li and Changyin Wu, date: 07/20/2014, view: 6489, Q&A: 0
In this protocol, we describe a simple and efficient method for meiotic chromosome spread preparation in rice pollen mother cells. Meiotic chromosome preparation by spreading itself is an important technique for plant cytogenetics (Higgins et al., 2004; Chelysheva et al., 2012; Wang et al., 2009); furthermore, it is a ...
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Stem Cell

Coculture between hMADS and Mouse Adult CM Authors:  Florence Figeac, Adrien Acquistapace, Olivier Le Coz, Pierre François Lesault and Anne-Marie Rodriguez, date: 07/20/2014, view: 6310, Q&A: 0
Heart failure occurring after acute myocardial infarction (MI) is among the main causes of death in western countries. Cell therapies, particularly those based on mesenchymal stem cells (MSC), represent one of the most promising approaches to repair damaged heart tissues. Several reports have provided evidences that injection of mesenchymal stem ...
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Mesenchymal Stem Cell (MSC) Aggregate Formation in vivo Authors:  Thomas J. Bartosh and Joni H. Ylostalo, date: 07/20/2014, view: 7852, Q&A: 0
Human mesenchymal stem/progenitor cells (MSCs) isolated from various adult tissues show remarkable therapeutic potential and are being employed in clinical trials for the treatment of numerous diseases (Prockop et al., 2010). While routes of cell administration vary, profound beneficial effects of MSCs in animal models have been observed ...
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