• Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011
Image Credit: BioProtoc.3702

Past Issues in 2020

Volume: 10 Issue: 15

Biophysics

Expression and Purification of Functionally Active Serotonin 5-HT2A Receptor in Insect Cells Using Low-titer Viral Stock Authors:  Sukanya Mozumder, Gopa Mahesh, Krishnamoorthi Srinivasan, Jayati Sengupta and Sujoy Mukherjee, date: 08/05/2020, view: 687, Q&A: 0
The serotonin 5-HT2A receptor (5-HT2AR) is a member of the GPCR family that is important for various neurological functions and whose dysregulation causes many mental health disorders. Structural investigations of 5-HT2AR require the production of functionally active receptors expressed from eukaryotic cell ...
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Cancer Biology

SMART (Single Molecule Analysis of Resection Tracks) Technique for Assessing DNA end-Resection in Response to DNA Damage Authors:  Angela Altieri, Milena Dell’Aquila, Francesca Pentimalli, Antonio Giordano and Alfano Luigi, date: 08/05/2020, view: 776, Q&A: 0
DNA double strand breaks (DSBs) are among the most toxic lesions affecting genome integrity. DSBs are mainly repaired through non-homologous end joining (NHEJ) and homologous recombination (HR). A crucial step of the HR process is the generation, through DNA end-resection, of a long 3′ single-strand DNA stretch, necessary to prime DNA synthesis ...
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Cell Biology

Measuring Intracellular Vesicle Density and Dispersion Using Fluorescence Microscopy and ImageJ/FIJI Authors:  Natália Fernanda do Couto, Thamires Queiroz-Oliveira, Maria Fátima Horta, Thiago Castro-Gomes and Luciana Oliveira Andrade, date: 08/05/2020, view: 1167, Q&A: 0
Cell signalling, cell secretion, and plasma membrane repair are processes that critically rely on intracellular vesicles, important components of the endocytic and secretory pathways. More specifically, the strategic distribution of intracellular vesicles is important for diverse cellular processes. The method presented here is a simple, ...
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Buoyant Density Fractionation of Small Extracellular Vesicle Sub-populations Derived from Mammalian Cells Authors:  Morayma M. Temoche-Diaz, Matthew J. Shurtleff and Randy Schekman, date: 08/05/2020, view: 647, Q&A: 0
Small extracellular vesicles (sEVs) encompass a variety of distinct vesicles that are secreted to the extracellular space. Many methodologies currently used for EV isolation (e.g., differential ultracentrifugation concluding in a high-speed pellet, precipitation by macromolecular crowding agents or size excusion chromatography–SEC) do not ...
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A Quantitative Single-cell Flow Cytometry Assay for Retrograde Membrane Trafficking Using Engineered Cholera Toxin Authors:  Mariska Simpson, Wayne I. Lencer and Phi Luong, date: 08/05/2020, view: 704, Q&A: 0
The organization and distribution of proteins, lipids, and nucleic acids in eukaryotic cells is an essential process for cell function. Retrograde trafficking from the plasma membrane to the Golgi and endoplasmic reticulum can greatly modify cell membrane composition and intracellular protein dynamics, and thus typifies a key sorting step. ...
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Developmental Biology

Mechanical Tissue Compression and Whole-mount Imaging at Single Cell Resolution for Developing Murine Epididymal Tubules Author:  Tsuyoshi Hirashima, date: 08/05/2020, view: 748, Q&A: 0
Cells inside the body are subjected to various mechanical stress, such as stretch or compression provided by surrounding cells, shear stresses by blood or lymph flows, and normal stresses by luminal liquids. Force loading to the biological tissues is a fundamental method to better understand cellular responses to such mechanical stimuli. There ...
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Immunology

Flow Cytometry of CD14, VDR, Cyp27 and Cyp24 and TLR4 in U937 Cells Authors:  Jacqueline Ferritto Rebello, Rodrigo Barbosa de Oliveira Brito, Caren Cristina Grabulosa, Rosa Maria Affonso Moyses, Rosilene Motta Elias and Maria Aparecida Dalboni, date: 08/05/2020, view: 630, Q&A: 0
Chronic Kidney Disease (CKD) patients present a micro inflammation state due to failure renal function. The calcitriol has been described as an anti-inflammatory factor that might modulates the inflammatory response in CKD patients. However, these patients have deficiency of Calcitriol due to failure renal function. But, synthesis of this vitamin ...
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Microbiology

High-level Production of Recombinant Membrane Proteins Using the Engineered Escherichia coli Strains SuptoxD and SuptoxR Authors:  Myrsini Michou, Dafni C. Delivoria and Georgios Skretas, date: 08/05/2020, view: 725, Q&A: 0
We have previously described the development of two specialized Escherichia coli strains for high-level recombinant membrane protein (MP) production. These engineered strains, termed SuptoxD and SuptoxR, are capable of suppressing the cytotoxicity caused by MP overexpression and of producing greatly enhanced MP yields. Here, we present a ...
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Paper Lateral Flow Biosensor for Nodavirus Reverse Transcribed RNA Detection Authors:  Dimitra K. Toubanaki and Evdokia Karagouni, date: 08/05/2020, view: 421, Q&A: 0
Paper nanobiosensors have been established as an excellent platform for analysis of veterinary and human pathogens causing various diseases. Especially, lateral flow assays or biosensors ideal for sensitive, rapid, robust and accurate analysis in laboratory setups and on-site analysis. Viral RNA detection is of great importance for public health ...
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Molecular Biology

A Method for User-defined Mutagenesis by Integrating Oligo Pool Synthesis Technology with Nicking Mutagenesis Authors:  Paul J. Steiner, Zachary T Baumer and Timothy A. Whitehead, date: 08/05/2020, view: 831, Q&A: 0
Saturation mutagenesis is a fundamental enabling technology for protein engineering and epitope mapping. Nicking mutagenesis (NM) allows the user to rapidly construct libraries of all possible single mutations in a target protein sequence from plasmid DNA in a one-pot procedure. Briefly, one strand of the plasmid DNA is degraded using a nicking ...
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Neuroscience

Multiple Simultaneous Acute Stresses in Mice: Single or Repeated Induction Authors:  Rachael E. Hokenson, Mikko Oijala, Annabel K. Short, Jessica L. Bolton, Yuncai Chen, Jenny Molet, Pamela M. Maras, Tallie Z. Baram and Gyorgy Lur, date: 08/05/2020, view: 805, Q&A: 0
Stress is crucial to the survival of an organism, but excessive stress can lead to psychological disorders including depression, anxiety, substance abuse, and suicidality. The prevailing notion is that chronic stress promotes adverse outcomes on brain and body health, whereas acute stressors are generally benign. Notably, acute events such mass ...
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Generation of Functional Mouse Hippocampal Neurons Authors:  Francesco Tomassoni-Ardori, Zhenyi Hong, Gianluca Fulgenzi and Lino Tessarollo, date: 08/05/2020, view: 897, Q&A: 0
Primary culture of mouse hippocampal neurons is a very useful in vitro model for studying neuronal development, axonal and dendritic morphology, synaptic functions, and many other neuronal features. Here we describe a step-by-step process of generating primary neurons from mouse embryonic hippocampi (E17.5/E18.5). Hippocampal neurons ...
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Development of the Deoxycorticosterone Acetate (DOCA)-salt Hypertensive Rat Model Authors:  Nayara Pestana-Oliveira, David B. Nahey, Tim Johnson and John P. Collister, date: 08/05/2020, view: 402, Q&A: 0
According to the World Health Organization (WHO), nearly 1.13 billion people worldwide have hypertension, a major factor responsible for premature death globally. The inherent multifactorial nature of hypertension makes its study difficult since the chronic rise in blood pressure depends on the intricate connection between dietary, genetic ...
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In vivo Blood-brain Barrier Permeability Assays Using Clostridium perfringens Epsilon Toxin Authors:  Michael R. Mazzucco, Timothy Vartanian and Jennifer R. Linden, date: 08/05/2020, view: 534, Q&A: 0
In order for the brain to function properly, a carefully orchestrated homeostasis must be maintained. To help regulate this delicate balance, the brain has developed a highly selective blood-brain barrier (BBB). Under normal conditions, the BBB excludes harmful blood-borne material from the brain parenchyma. However, numerous neuropathological ...
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Plant Science

The ATPase Activity of Escherichia coli Expressed AAA+-ATPase Protein Authors:  Amita Kaundal, Vemanna S. Ramu and Kirankumar S. Mysore, date: 08/05/2020, view: 601, Q&A: 0
ATPases are the enzymes that breakdown ATP to ADP and release inorganic phosphate (Pi). Here we provide a detailed protocol to determine the ATPase activity of a recombinant AAA+-ATPase protein (GENERAL CONTROL NON-REPRESSIBLE-4 [GCN4]) by spectrophotometric absorption at 360 nm to measure the accumulated inorganic phosphate. In ...
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