• 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

Wheat Germ Agglutinin (WGA)-SDS-PAGE: A Novel Method for the Detection of O-GlcNAc-modified Proteins by Lectin Affinity Gel Electrophoresis Authors:  Ko Fujioka, Yuji Kubota and Mutsuhiro Takekawa, date: 12/05/2018, view: 112, Q&A: 0
Diverse cytoplasmic and nuclear proteins dynamically change their molecular functions by O-linked β-N-acetylglucosamine (O-GlcNAc) modification on serine and/or threonine residues. Evaluation of the O-GlcNAcylation level of a specific protein, however, needs multiple and time-consuming steps if using conventional methods (e.g., ...
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Purification of Globular Actin from Rabbit Muscle and Pyrene Fluorescent Assays to Investigate Actin Dynamics in vitro Authors:  He Sun, Yuanyuan Luo and Yansong Miao, date: 12/05/2018, view: 81, Q&A: 0
Pyrene Fluorescent Assay is established to monitor the dynamic actin nucleation, elongation, capping and disassembly in vitro. This technique provides an easy handle procedure and straightforward visual data analysis. By coupling actin purification and polymerization assays in this protocol, the readers could quickly get the affordable ...
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In vitro Membrane Interaction and Liposome Fusion Assays Using Recombinant Hepatitis C Virus Envelope Protein E2 Authors:  Belén Yélamos, Mar Rodríguez-Rodríguez, Francisco Gavilanes and Julián Gómez-Gutiérrez, date: 12/05/2018, view: 54, Q&A: 0
In order to study the mechanism underlying the Hepatitis C Virus (HCV) fusion process we have performed assays using phospholipid liposomes and a truncated form of E2 protein, E2661 (amino acids 384-661 of the HCV polyprotein) lacking the transmembrane region. E2661 has been previously generated by using the baculovirus ...
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Cancer Biology

Cluster Analysis of Endogenous HER2 and HER3 Receptors in SKBR3 Cells Authors:  Selene K. Roberts, Michael Hirsch, Alexandra McStea, Laura C. Zanetti-Domingues, David T. Clarke, Jeroen Claus, Peter J. Parker, Lin Wang and Marisa L. Martin-Fernande, date: 12/05/2018, view: 127, Q&A: 0
The Human Epidermal Growth Factor Receptor (HER) family of receptor tyrosine kinases consists of four, single pass, transmembrane receptor homologs (HER1-4) that act to regulate many critical processes in normal and tumor cells. HER2 is overexpressed in many tumors, and the deregulated proliferation of cancerous cells is driven by cooperation with ...
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Cell Biology

Blinded Visual Scoring of Images Using the Freely-available Software Blinder Authors:  Steven D. Cothren, Joel N. Meyer and Jessica H. Hartman, date: 12/05/2018, view: 94, Q&A: 0
In nearly all subfields of biomedical sciences, there are phenotypes that are currently classified by expert visual scoring. In research applications, these classifications require the experimenter to be blinded to the treatment group in order to avoid unintentional bias in scoring. Currently, many labs either use laborious and tedious methods to ...
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Developmental Biology

Notochord Injury Assays that Stimulate Transcriptional Responses in Zebrafish Larvae Authors:  Zhiqiang Zeng, Juan C Lopez-Baez, Laura Lleras-Forero, Hannah Brunsdon, Cameron Wyatt, Witold Rybski, Nicholas D Hastie, Stefan Schulte-Merker and E Elizabeth Patton, date: 12/05/2018, view: 125, Q&A: 0
Zebrafish have become an increasingly important model organism in the field of wound healing and regenerative medicine, due to their high regenerative capacity coupled with high-resolution imaging in living animals. In a recent study, we described multiple physical and chemical methods to induce notochord injury that led to highly specific ...
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Immunology

Flow Cytometry Assay for Recycling of LFA-1 in T-lymphocytes Authors:  Katarzyna Potrzebowska, Janne Lehtonen, Malin Samuelsson and Lena Svensson, date: 12/05/2018, view: 101, Q&A: 0
To enable cells to move forward, cell surface integrins are internalized into an endosomal compartment and subsequently intracellularly transported to be re-exposed at a new site on the cell membrane. Leukocytes are the fastest migrating cell type in the human body, which express the leukocyte-specific integrin LFA-1. Here, we describe a flow ...
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Integrin Dependent RhoB Activation Assay Using Leukocytes Authors:  Janne Lehtonen, Katarzyna Potrzebowska, Malin Samuelsson and Lena Svensson, date: 12/05/2018, view: 58, Q&A: 0
To be able to migrate, leukocyte needs to re-use its adhesion molecules to move forward. These adhesion molecules are called integrins and are intracellularly transported via endocytosis and exocytosis in order to translocate to a new site on the cell membrane. The intracellular transportation is regulated by different small GTPases including ...
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Ultrasound Guided Intra-thymic Injection to Track Recent Thymic Emigrants and Investigate T Cell Development Authors:  Haiguang Wang, David L. Owen, Lily J. Qian, Laura B. Chopp, Michael A. Farrar and Kristin A. Hogquist, date: 12/05/2018, view: 94, Q&A: 0
To track recent thymic emigrants (RTEs) or study T cell development in the thymus, intra-thymic injection of a cellular tag or precursor cells for various T cell lineages is often desired. However, the traditional surgical approach to expose the thymus for intra-thymic injection is time-consuming and can cause a high level of pain and stress to ...
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Microbiology

Field Inoculation and Classification of Maize Ear Rot Caused by Fusarium verticillioides Authors:  Chaopei Dong, Yabin Wu, Jingyang Gao, Zijian Zhou, Cong Mu, Peipei Ma, Jiafa Chen and Jianyu Wu, date: 12/05/2018, view: 70, Q&A: 0
Maize ear rot is a worldwide fungal disease mainly caused by Fusarium verticillioides and Fusarium graminearum. Maize planted in the field was inoculated with Fusarium verticillioides at the filling stage, 15 days after pollination. Two milliliters of spore suspension with a concentration of 5 x 106/ml was ...
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Quantitative Transformation Efficiency Assay for Bacillus subtilis Authors:  Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 153, Q&A: 0
Bacillus subtilis (B. subtilis) is a model Gram-positive organism used to study a variety of physiological states and stress responses, one of which is the development of competence. Competence refers to the physiological state of a cell which allows it to be transformed naturally. Through induction of competence, the efficiency ...
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An in vitro Microscopy-based Assay for Microtubule-binding and Microtubule-crosslinking by Budding Yeast Microtubule-associated Protein Authors:  Yili Zhu, Weimin Tan and Wei-Lih Lee, date: 12/05/2018, view: 212, Q&A: 0
In this protocol, we describe a simple microscopy-based method to assess the interaction of a microtubule-associated protein (MAP) with microtubules. The interaction between MAP and microtubules is typically assessed by a co-sedimentation assay, which measures the amount of MAP that co-pellets with microtubules by centrifugation, followed by ...
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Plant Science

Genome-wide Estimation of Evolutionary Distance and Phylogenetic Analysis of Homologous Genes Authors:  Meixia Zhao, Biao Zhang, Jianxin Ma and Damon Lisch, date: 12/05/2018, view: 123, Q&A: 0
Homologous genes, including paralogs and orthologs, are genes that share sequence homologies within or between different species. Homologous genes originate from a common origin through speciation, genetic duplication or horizontal gene transfer. Estimation of the sequence divergence of homologous genes help us to understand divergence time, which ...
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Using Arabidopsis Mesophyll Protoplasts to Study Unfolded Protein Response Signaling Authors:  Yan Bao and Diane C. Bassham, date: 12/05/2018, view: 154, Q&A: 0
Various environmental stresses or artificial reagents can trigger unfolded protein accumulation in the endoplasmic reticulum (ER) due to the folding capacity of the ER being exceeded. This is termed ER stress, and triggers the unfolded protein response (UPR). Assays for activation of the UPR in plants include Tunicamycin (Tm)- or dithiothreitol ...
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