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Editors' Choice
Editors' Choice corresponds to recently published bio-protocols (past one year) that chosen by Bio-protocol Associate Editors based on the protocols' quality, novelty, or potential impact.
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Labeling Aversive Memory Trace in Mouse Using a Doxycycline-inducible Expression System
Authors:  Jianyang Du and Erin E. Koffman, date: 10/20/2017, view: 924, Q&A: 0
A memory trace, also known as a memory engram, is theorized to be a mechanism for physical memory storage in the brain (Silva et al., 2009; Josselyn, 2010) and memory trace is associated with a specific population of neurons (Liu et al., 2012; Ramirez et al., 2013). Labeling and stimulating those neurons will activate ...
Isolation and Purification of Schwann Cells from Spinal Nerves of Neonatal Rat
Authors:  Jinkun Wen, Dandan Tan, Lixia Li and Jiasong Guo, date: 10/20/2017, view: 956, Q&A: 0
Primary cultured Schwann cells (SCs) are widely used in the investigation of the biology of SC and are important seed cells for neural tissue engineering. Here, we describe a novel protocol for harvesting primary cultured SCs from neonatal Sprague-Dawley (SD) rats. In the present protocol, dissociated SCs are isolated from the spinal nerves of ...
Large-scale Maize Seedling Infection with Exserohilum turcicum in the Greenhouse
Authors:  Ping Yang, Gerhard Herren, Simon G. Krattinger and Beat Keller, date: 10/05/2017, view: 970, Q&A: 0
Northern corn leaf blight (NCLB) is a serious foliar disease of maize (Zea mays) worldwide and breeding for resistance is of primary importance for maize crop protection. Phenotyping for NCLB resistance is well established in the field, but such experiments depend on suitable environmental conditions and are seasonal. Here we describe a ...
Accurate, Streamlined Analysis of mRNA Translation by Sucrose Gradient Fractionation
Authors:  Soufiane Aboulhouda, Rachael Di Santo, Gabriel Therizols and David Weinberg, date: 10/05/2017, view: 1105, Q&A: 0
The efficiency with which proteins are produced from mRNA molecules can vary widely across transcripts, cell types, and cellular states. Methods that accurately assay the translational efficiency of mRNAs are critical to gaining a mechanistic understanding of post-transcriptional gene regulation. One way to measure translational efficiency is to ...
Peroxisome Motility Measurement and Quantification Assay
Authors:  Jeremy Metz, Inês G. Castro and Michael Schrader, date: 09/05/2017, view: 1089, Q&A: 0
Organelle movement, distribution and interaction contribute to the organisation of the eukaryotic cell. Peroxisomes are multifunctional organelles which contribute to cellular lipid metabolism and ROS homeostasis. They distribute uniformly in mammalian cells and move along microtubules via kinesin and dynein motors. Their metabolic cooperation ...
An ex vivo Perifusion Method for Quantitative Determination of Neuropeptide Release from Mouse Hypothalamic Explants
Authors:  Ophélia Le Thuc, Jacques Noël and Carole Rovère, date: 08/20/2017, view: 1095, Q&A: 0
The hypothalamus is a primary brain area which, in mammals, regulates several physiological functions that are all related to maintaining general homeostasis, by linking the central nervous system (CNS) and the periphery. The hypothalamus itself can be considered an endocrine brain region of some sort as it hosts in its different nuclei several ...
Protocol for HeLa Cells Infection with Escherichia coli Strains Producing Colibactin and Quantification of the Induced DNA-damage
Strains of Escherichia coli bearing the pks genomic island synthesize the genotoxin colibactin. Exposure of eukaryotic cells to E. coli producing colibactin induces DNA damages, ultimately leading to cell cycle arrest, senescence and death. Here we describe a simple method to demonstrate the genotoxicity of bacteria ...
Liposome Disruption Assay to Examine Lytic Properties of Biomolecules
Authors:  John R. Jimah, Paul H. Schlesinger and Niraj H. Tolia, date: 08/05/2017, view: 1545, Q&A: 0
Proteins may have three dimensional structural or amino acid features that suggest a role in targeting and disrupting lipids within cell membranes. It is often necessary to experimentally investigate if these proteins and biomolecules are able to disrupt membranes in order to conclusively characterize the function of these biomolecules. Here, we ...
Membrane Lipid Screen to Identify Molecular Targets of Biomolecules
Authors:  John R. Jimah, Paul H. Schlesinger and Niraj H. Tolia, date: 08/05/2017, view: 1330, Q&A: 0
Proteins that bind to and disrupt cell membranes may target specific phospholipids. Here we describe a protocol to identify the lipid targets of proteins and biomolecules. First, we describe a screen to identify lipids in membranes that are specifically bound by the biomolecule of interest. Second, we describe a method for determining if the ...
Bioelectrospray Methodology for Dissection of the Host-pathogen Interaction in Human Tuberculosis
Authors:  Liku B Tezera, Magdalena K Bielecka and Paul T Elkington, date: 07/20/2017, view: 1431, Q&A: 0
Standard cell culture models have been used to investigate disease pathology and to test new therapies for over fifty years. However, these model systems have often failed to mimic the changes occurring within three-dimensional (3-D) space where pathology occurs in vivo. To truthfully represent this, an emerging paradigm in biology is the ...