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    Protocols in Current Issue
    Microplastic Extraction from Marine Vertebrate Digestive Tracts, Regurgitates and Scats: A Protocol for Researchers from All Experience Levels
    Authors:  Amy L. Lusher and Gema Hernandez-Milian, date: 11/20/2018, view: 588, Q&A: 0
    [Abstract] It is essential to provide a protocol for the separation and identification of microplastics in marine vertebrates (mammals, birds, turtles and fish) that is easy to follow and adaptable depending on research infrastructure. Digesting organic material is an effective way to analyze samples for microplastics. Presented here is an optimized protocol ...
    Studying the Mechanisms of Developmental Vocal Learning and Adult Vocal Performance in Zebra Finches through Lentiviral Injection
    Authors:  Zhimin Shi, Ofer Tchernichovski and XiaoChing Li, date: 09/05/2018, view: 473, Q&A: 0
    [Abstract] Here we provide a detailed step-by-step protocol for using lentivirus to manipulate miRNA expression in Area X of juvenile zebra finches and for analyzing the consequences on song learning and song performance. This protocol has four parts: 1) making the lentiviral construct to overexpress miRNA miR-9; 2) packaging the lentiviral vector; 3) ...
    Xenopus laevis Oocytes Preparation for in-Cell EPR Spectroscopy
    Authors:  Laura John and Malte Drescher, date: 04/05/2018, view: 1286, Q&A: 0
    [Abstract] One of the most exciting perspectives for studying bio-macromolecules comes from the emerging field of in-cell spectroscopy, which enables to determine the structure and dynamics of bio-macromolecules in the cell. In-cell electron paramagnetic resonance (EPR) spectroscopy in combination with micro-injection of bio-macromolecules into Xenopus ...
    Boron Uptake Assay in Xenopus laevis Oocytes
    Authors:  Sheliang Wang, Namiki Mitani-Ueno and Junpei Takano, date: 03/05/2018, view: 1429, Q&A: 0
    [Abstract] Boron (B) is essential for plant growth and taken up by plant roots as boric acid. Under B limitation, B uptake and translocation in plants are dependent on the boric acid channels located in the plasma membrane. Xenopus leavis oocyte is a reliable heterologous expression system to characterize transport activities of boric acid channels ...
    In vitro Engineered DNA-binding Molecule-mediated Chromatin Immunoprecipitation (in vitro enChIP) Using CRISPR Ribonucleoproteins in Combination with Next-generation Sequencing (in vitro enChIP-Seq) for the Identification of Chromosomal Interactions
    Authors:  Toshitsugu Fujita and Hodaka Fujii, date: 11/20/2017, view: 2876, Q&A: 0
    [Abstract] We have developed locus-specific chromatin immunoprecipitation (locus-specific ChIP) technologies consisting of insertional ChIP (iChIP) and engineered DNA-binding molecule-mediated ChIP (enChIP). Locus-specific ChIP is a method to isolate a genomic region of interest from cells while it also identifies what binds to this region using mass ...
    Preparation of the Partially Methylated Alditol Acetates Derived from CS Tetrasaccharides Containing Galactose for the Gas Chromatography/Mass Spectrometry Analysis
    Authors:  Kyohei Higashi and Toshihiko Toida, date: 11/05/2017, view: 2397, Q&A: 0
    [Abstract] Chondroitin sulfate (CS), a member of the glycosaminoglycan (GAG) family of carbohydrates, is composed of linear, sulfated repeating disaccharide sequences of N-acetyl-D-galactosamine (GalNAc) and glucuronic acid (GlcA). Recently, a keratan sulfate (KS) disaccharide [GlcNAc6S(β1-3)Galactose(β1-]-branched CS-E was identified ...
    Uptake Assays in Xenopus laevis Oocytes Using Liquid Chromatography-mass Spectrometry to Detect Transport Activity
    [Abstract] Xenopus laevis oocytes are a widely used model system for characterization of heterologously expressed secondary active transporters. Historically, researchers have relied on detecting transport activity by measuring accumulation of radiolabeled substrates by scintillation counting or of fluorescently labelled substrates by ...
    Isolation of Keratan Sulfate Disaccharide-branched Chondroitin Sulfate E from Mactra chinensis
    Authors:  Kyohei Higashi and Toshihiko Toida, date: 08/05/2017, view: 2291, Q&A: 0
    [Abstract] Glycosaminoglycans (GAGs) including chondroitin sulfate (CS), dermatan sulfate (DS), heparin (HP), heparan sulfate (HS) and keratan sulfate (KS) are linear, sulfated repeating disaccharide sequences containing hexosamine and uronic acid (or galactose in the case of KS). Recently, a keratan sulfate (KS) disaccharide [GlcNAc6S ...
    Spinal Cord Preparation from Adult Red-eared Turtles for Electrophysiological Recordings during Motor Activity
    Authors:  Peter C Petersen and Rune W Berg , date: 07/05/2017, view: 4256, Q&A: 0
    [Abstract] Although it is known that the generation of movements is performed to a large extent in neuronal circuits located in the spinal cord, the involved mechanisms are still unclear. The turtle as a model system for investigating spinal motor activity has advantages, which far exceeds those of model systems using other animals. The high resistance to ...
    Expression and Analysis of Flow-regulated Ion Channels in Xenopus Oocytes
    Authors:  Shujie Shi and Marcelo D. Carattino, date: 04/20/2017, view: 3691, Q&A: 0
    [Abstract] Mechanically-gated ion channels play key roles in mechanotransduction, a process that translates physical forces into biological signals. Epithelial and endothelial cells are exposed to laminar shear stress (LSS), a tangential force exerted by flowing fluids against the wall of vessels and epithelia. The protocol outlined herein has been used to ...



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