Volume: 5 Issue: 20 —BIO-PROTOCOL

Cancer Biology

Rat Aortic Ring Model to Assay Angiogenesis ex vivo Authors: Isabelle Ernens, Bénédicte Lenoir, Yvan Devaux and Daniel R. Wagner, date: 10/20/2015, view: 7414, Q&A: 0

Angiogenesis is a multifactorial event which requires the migration, proliferation, differentiation and structure rearrangement of endothelial cells. This angiogenic process has been commonly studied using in vitro assays such as Boyden chamber assay, wound healing assay and tube formation assay. These assays mainly use monolayers of ...

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Cell Biology

In vivo Fluorescein Isothiocyanate-dextran (FD4) Permeability Assay Authors: Carey L Watson, Maxime M Mahe and Michael A. Helmrath, date: 10/20/2015, view: 15028, Q&A: 1

Using pluripotent stem cells, it is now becoming possible to develop tissue models of organ systems within the body. These organs will allow for the study of organ function, physiology, embryology, and even pathologic processes. Recently, our group developed a model of human small intestine developed from human pluripotent stem cells which when ...

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Intravenous Tomato Lectin Injection to Assess Functional Vasculature Authors: Carey L Watson, Maxime M Mahe and Michael A. Helmrath, date: 10/20/2015, view: 8522, Q&A: 1

Pluripotent stem cells have recently allowed for the development of tissue models for the various organ systems within the body. These models allow scientists to study organ function, physiology, embryology, and even pathologic processes. Studies on tissue can be done in vitro and/or transplanted into animal models for studies in vivo ...

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Immunology

Coupling of HIV-1 gp120-derived Core Protein to Paramagnetic Beads and Adsorption Assays Authors: Jidnyasa Ingale and Richard T Wyatt, date: 10/05/2015, view: 5097, Q&A: 0

Analysis of the functional activity in polyclonal serum following immunization of a complex protein or glycoprotein immunogen is a very important but tedious process. Fine mapping of epitope-specific antibodies is difficult when they are elicited at relatively low levels. In our recent study focused on developing an HIV-1 vaccine, we immunized ...

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Kinetic Analysis of Monoclonal Antibody Binding to HIV-1 gp120-derived Hyperglycosylated Cores Authors: Jidnyasa Ingale and Richard T Wyatt, date: 10/05/2015, view: 5859, Q&A: 0

Kinetic analysis of antibodies is one of the important studies for characterization of antibodies and screening of ligands. In our recent study (Ingale et al., 2014), we compared the antigenic profiles and binding characteristics of four HIV-1 envelope glycoprotein (Env) core immunogens using multiple monoclonal antibodies by Bio-Layer ...

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Isolation and Culture of Human Endometrial Epithelial Cells and Stromal Fibroblasts Authors: Joseph C. Chen and Nadia R. Roan, date: 10/20/2015, view: 13101, Q&A: 0

Purification and culture of endometrial epithelial cells (eEC) and stromal fibroblasts (eSF) from endometrial biopsies allows for downstream cell-specific in vitro studies. The utility of this protocol is the ease with which cells are purified without contamination from unwanted cell types, and the ability to use patient-paired eEC and ...

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Collagen-induced Arthritis: A Model for Murine Autoimmune Arthritis Authors: Kathryn M. Pietrosimone, Mengyao Jin, Brad Poston and Peng Liu, date: 10/20/2015, view: 9421, Q&A: 0

Collagen-induced arthritis (CIA) is a common autoimmune animal model used to study rheumatoid arthritis (RA). The development of CIA involves infiltration of macrophages and neutrophils into the joint, as well as T and B cell responses to type II collagen. In murine CIA, genetically susceptible mice (DBA/1J) are immunized with a type II bovine ...

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Immune Cell Isolation from Mouse Femur Bone Marrow Authors: Xiaoyu Liu and Ning Quan, date: 10/20/2015, view: 16026, Q&A: 0

The bone marrow is the site of hematopoiesis and contains mixed population of blood cells including erythrocytes, granulocytes, monocytes, dendritic cells, lymphocytes and hematopoietic stem cells. The following protocol provides a simple and fast method for isolation of bone marrow immune cells (no erythrocytes) from mouse femurs with a yield of ...

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Microbiology

Product Analysis of Starch Active Enzymes by TLC Authors: Darrell Cockburn and Nicole Koropatkin, date: 10/20/2015, view: 8488, Q&A: 0

Thin layer chromatography (TLC) is a useful technique for detecting the presence of monosaccharides through to oligosaccharides, though it needs to be optimized for the specific sugars that are analyzed. Here we present a method for visualizing the reaction product(s) of starch active enzymes, which can contain α-1, 4 linked and α-1, 6 linked ...

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Purification of a Protein Exhibiting Isoleucine 2-epimerase Activity from Lactobacillus otakiensis JCM 15040 Authors: Yuta Mutaguchi and Toshihisa Ohshima, date: 10/20/2015, view: 5295, Q&A: 0

Prominent accumulation of D-leucine, D-allo-isoleucine and D-valine was observed in the culture medium of the heterofermentative bacterial species, Lactobacillus otakiensis (L. otakiensis) JCM 15040. The racemase enzyme that resulted in this accumulation, isoleucine 2-epimerase, was purified from the bacterial cells. ...

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Plant Science

Mitochondrial RNA Transcript Analysis Assay of Arabidopsis Leaf Tissues Authors: Etienne Delannoy, Andéol Falcon de Longevialle and Catherine Colas des Francs-Small, date: 10/20/2015, view: 6092, Q&A: 0

This qPCR-based assay provides an overview of the expression levels of all mitochondrial transcripts (mRNAs and rRNAs) as well as splicing efficiency in Arabidopsis. It was developed before RNAseq techniques were widely used (de Longevialle et al., 2007), but is nevertheless still useful as it is cheaper to run and the analysis ...

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Isolation and Characterization Procedure for Indole Alkaloids from the Marquesan Plant Rauvolfia Nukuhivensis Authors: Nicolas J. Martin, Maël Nicolas, Gaël Lecellier and Phila Raharivelomanana, date: 10/20/2015, view: 7933, Q&A: 0

A plethora of natural products, mostly secondary metabolites, are isolated and purified from many different organisms, like plants, fungi, algae, marine invertebrates, etc. The extraction procedure is specific to each organism, but some guidelines are usually followed for any purification procedure regarding targeted metabolites, such as ...

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Hyaloperonospora arabidopsidis (Downy Mildew) Infection Assay in Arabidopsis Authors: Shuta Asai, Ken Shirasu and Jonathan D. G. Jones, date: 10/20/2015, view: 11357, Q&A: 2

Hyaloperonospora arabidopsidis (Hpa; formerly Peronospora parasitica or Hyaloperonospora parasitica) is an oomycete downy mildew pathogen of the model plant Arabidopsis. The pathosystem between Arabidopsis and Hpa has been extensively used to study host/pathogen co-evolution (Coates ...

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Two-photon Photoactivation to Measure Histone Exchange Dynamics in Plant Root Cells Authors: Stefanie Rosa and Peter Shaw, date: 10/20/2015, view: 5412, Q&A: 0

Chromatin-binding proteins play a crucial role in chromatin structure and gene expression. Direct binding of chromatin proteins both maintains and regulates transcriptional states. It is therefore important to study the binding properties of these proteins in vivo within the natural environment of the nucleus. Photobleaching, ...

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Vacuole Structure Analysis during Cell Death Subsequent to Application of Erwinia carotovora Culture Filtrates to Cell Cultures of Nicotiana tabacum Authors: Yumi Hirakawa, Seiichiro Hasezawa and Takumi Higaki, date: 10/20/2015, view: 5205, Q&A: 0

We recently established an experimental model system for efficient defense-related cell death using tobacco BY-2 cultured cells treated with culture filtrates of the pathogenic bacterium Erwinia carotovora (E. carotovora) (Hirakawa et al., 2015). Applying this experimental system to transgenic BY-2 cells stably ...

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Isolation of Triterpenes from Propolis (Bee Glue) Authors: Muhammad N. Kardar and Veronique Seidel, date: 10/20/2015, view: 9805, Q&A: 1

Propolis (bee glue) is a natural substance produced by bees upon collection of mainly plant resins. Bees use it as antiseptic sealing agent between honeycombs and to preserve the hive from external contamination. Numerous scientific studies have been published on the biological properties of propolis including its anti-inflammatory, anti-oxidant, ...

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Stem Cell

Clonal Culture of Mouse Liver Progenitor Cells Author: Naoki Tanimizu, date: 10/20/2015, view: 8364, Q&A: 0

Liver stem/progenitor cells (LPCs) are defined as bipotential cells differentiating into both hepatocytes and cholangiocytes. For analyzing their differentiation potential, clonal culture has been used for LPCs isolated by a cell sorter. In addition, we can use the culture to assess functions of target genes on differentiation potential of LPCs. ...

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Past Issues in 2015

Volume: 5 Issue: 20

Cancer Biology

Cell Biology

Immunology

Microbiology

Plant Science

Stem Cell