• 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

Separation and Visualization of Low Abundant Ubiquitylated Forms Authors:  Ramona Schuster, Tânia Simões, Fabian den Brave and Mafalda Escobar-Henriques, date: 11/20/2018, view: 449, Q&A: 0
In this protocol we describe the separation and visualization of ubiquitylated forms of the yeast mitofusin Fzo1 by Western blot. To this aim, we express HA-tagged Fzo1 in Saccharomyces cerevisiae, break the cells to extract a membrane-enriched fraction, solubilize the membranes using detergent and then specifically immunoprecipitate the ...
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Cancer Biology

A Functionally Robust Phenotypic Screen that Identifies Drug Resistance-associated Genes Using 3D Cell Culture Authors:  Sun-Young Lee and Mina J. Bissell, date: 11/20/2018, view: 463, Q&A: 0
Drug resistance is a major obstacle in cancer treatment: A case in point is the failure of cancer patients to respond to tyrosine kinase inhibitors (TKI) of EGFR, a receptor that is highly expressed in many cancers. Identification of the targets and delineation of mechanisms of drug resistance remain major challenges. Traditional pharmacological ...
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Cell Biology

In-vivo gp100-specific Cytotoxic CD8+ T Cell Killing Assay Authors:  Mahdia Benkhoucha, Nicolas Molnarfi, Elodie Belnoue, Madiha Derouazi and Patrice H Lalive, date: 11/20/2018, view: 358, Q&A: 0
Cytotoxic CD8+ T lymphocytes (CTLs) represent a crucial component of the adaptive immune system and play a prominent role in the anti-tumor immune responses of both mice and humans.

Cytotoxic CD8+ T cells are responsible for the lysis of cells expressing peptides associated with MHC class I molecules and derived ...
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Relative Quantitation of Polymerized Actin in Suspension Cells by Flow Cytometry Authors:  Mallory R. Kakley, Katrina B. Velle and Lillian K. Fritz-Laylin, date: 11/20/2018, view: 350, Q&A: 0
The amount of polymerized actin within a cell can vary widely due to natural processes and/or experimentally induced perturbations. We routinely use this protocol to measure relative polymerized actin content between cell populations by staining cells in suspension with fluorescent phalloidin, then measuring total cell fluorescence using flow ...
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Developmental Biology

Electron Microscopic Detection of Proteins and Protein Complexes in Mammalian Cells Using APEX-tagged, Conditionally Stable Nanobodies Authors:  Thomas E. Hall, James Rae, Nicholas Ariotti and Robert G. Parton, date: 11/20/2018, view: 268, Q&A: 0
The recent development of 3D electron microscopic techniques for cells and tissues has necessitated the development of new methods for the detection of proteins and protein-complexes in situ. The development of new genetic tags, such as the ascorbate peroxidase, APEX, for electron microscopic detection of tagged proteins has expanded the ...
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Mosaic Labeling and 3-Dimensional Morphological Analysis of Single Cells in the Zebrafish Left-right Organizer Authors:  Agnik Dasgupta, Andrew E. Jacob and Jeffrey D. Amack, date: 11/20/2018, view: 203, Q&A: 0
A transient epithelial structure called the left-right organizer (LRO) establishes left-right asymmetry in vertebrate embryos. Developmental defects that alter LRO formation result in left-right patterning errors that often lead to congenital heart malformations. However, little is known about mechanisms that regulate individual cell behaviors ...
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Environmental science

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: 590, Q&A: 0
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 ...
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Microbiology

Use of Gas Chromatography to Quantify Short Chain Fatty Acids in the Serum, Colonic Luminal Content and Feces of Mice Authors:  Willian Rodrigues Ribeiro, Marco Aurélio Ramirez Vinolo, Leandro Augusto Calixto and Caroline Marcantonio Ferreira, date: 11/20/2018, view: 275, Q&A: 0
Short-Chain Fatty Acids (SCFAs) are a product of the fermentation of resistant starches and dietary fibers by the gut microbiota. The most important SCFA are acetate (C2), propionate (C3) and butyrate (C4). These metabolites are formed and absorbed in the colon and then transported through the hepatic vein to the liver. SCFAs are more concentrated ...
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Neuroscience

Purification of Soluble Recombinant Human Tau Protein from Bacteria Using Double-tag Affinity Purification Authors:  Joseph McInnes, Lujia Zhou and Patrik Verstreken, date: 11/20/2018, view: 333, Q&A: 0
Dysfunction of the microtubule-associated protein Tau (encoded by the MAPT gene) has been implicated in more than twenty neurodegenerative diseases, including Alzheimer’s. As such, the physiological and disease-relevant functions of Tau have garnered great interest in the research community. One barrier hampering investigations into the ...
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A Modified Barnes Maze for Juvenile Rats Authors:  Nazanin Valibeigi, Daniel G. McHail, Carmen L. Kimball and Theodore C. Dumas, date: 11/20/2018, view: 231, Q&A: 0
To better understand neural codes for spatial navigation and to address cognitive development questions, the neurobiology of postnatal hippocampal development is receiving increased attention. While the Morris Water Maze is the gold standard for assessing hippocampal integrity, potential confounds can be difficult to control for in juvenile ...
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Papain-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry Authors:  Hanadie Yousef, Cathrin J Czupalla, Davis Lee, Eugene C Butcher and Tony Wyss-Coray, date: 11/20/2018, view: 240, Q&A: 0
Brain endothelial cells (BECs) form the integral component of the blood-brain barrier (BBB) which separates the systemic milieu from the brain parenchyma and protects the brain from pathogens and circulating factors. In order to study BEC biology, it was of particular interest to establish a method that enables researchers to investigate and ...
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Collagenase-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry Authors:  Cathrin J. Czupalla, Hanadie Yousef, Tony Wyss-Coray and Eugene C. Butcher, date: 11/20/2018, view: 231, Q&A: 0
The brain endothelium is a highly specialized vascular structure that maintains the activity and integrity of the central nervous system (CNS). Previous studies have reported that the integrity of the brain endothelium is compromised in a plethora of neuropathologies. Therefore, it is of particular interest to establish a method that enables ...
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Stem Cell

One-step Derivation of Functional Mesenchymal Stem Cells from Human Pluripotent Stem Cells Authors:  Yan Zhou, JinQi Liao, ChongZhou Fang, CuiPing Mo, GuangQian Zhou and Yonglun Luo, date: 11/20/2018, view: 231, Q&A: 0
Mesenchymal stem cells (MSCs) are invaluable cell sources for understanding stem cell biology and potential application in tissue engineering and regenerative medicine. The current issues of MSCs that demand to be further addressed are limited donors, tissue sources and limited capacity of ex vivo expansion. Here, we describe a simple and ...
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