• 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.3681

Past Issues in 2020

Volume: 10 Issue: 14

Biochemistry

Immunofluorescent Staining of Claudin-2 in Cultured Kidney Tubular Cells Authors:  Shaista Anwer and Katalin Szaszi, date: 07/20/2020, view: 606, Q&A: 0
Members of the claudin family of tight junction proteins regulate paracellular permeability and modulate cell signaling. During junction remodeling, these proteins are selectively inserted into or retrieved from the tight junctions, but the control and coordination of these processes remain incompletely understood. Visualization of claudins allows ...
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Biophysics

Production and Isolation of Magnetic Protein Crystals in HEK293T Cells Authors:  Thomas L. Li and Bianxiao Cui, date: 07/20/2020, view: 723, Q&A: 0
Advances in protein engineering have enabled the production of self-assembled protein crystals within living cells. Our recent publication demonstrates the production of ftn-PAK4, which is a ferritin-containing crystal that can mineralize iron and become magnetic when isolated. We have developed an optimized protocol for the production and ...
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Preparation and Characterization of Ginger Lipid-derived Nanoparticles for Colon-targeted siRNA Delivery Authors:  Junsik Sung, Chunhua Yang, James F. Collins and Didier Merlin, date: 07/20/2020, view: 523, Q&A: 0
Synthetic nanoparticle-based drug delivery system is widely known for its ability to increase the efficacy and specificity of loaded drugs, but it often suffers from relatively higher immunotoxicity and higher costs as compared to traditional drug formulations. Contrarily, plant-derived nanoparticles appear to be free from these limitations of ...
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Cancer Biology

A CRISPR Competition Assay to Identify Cancer Genetic Dependencies Authors:  Vishruth Girish and Jason M. Sheltzer, date: 07/20/2020, view: 1505, Q&A: 0
The CRISPR/Cas9 system is a powerful tool for genome editing, wherein the RNA-guided nuclease Cas9 can be directed to introduce double-stranded breaks (DSBs) at a targeted locus. In mammalian cells, these DSBs are typically repaired through error-prone processes, resulting in insertions or deletions (indels) at the targeted locus. Researchers can ...
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Developmental Biology

Isolation, Culture, and Differentiation of Primary Myoblasts Derived from Muscle Satellite Cells Authors:  Kun Ho Kim, Jiamin Qiu and Shihuan Kuang, date: 07/20/2020, view: 742, Q&A: 0
The skeletal muscle is key for body mobility and motor performance, but aging and diseases often lead to progressive loss of muscle mass due to wasting or degeneration of muscle cells. Muscle satellite cells (MuSCs) represent a population of tissue stem cells residing in the skeletal muscles and are responsible for homeostatic maintenance and ...
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In vitro Induction and Detection of Acrosomal Exocytosis in Human Spermatozoa Authors:  Shenae L. Cafe, Amanda L. Anderson and Brett Nixon, date: 07/20/2020, view: 522, Q&A: 0
The acrosome reaction is a highly regulated exocytotic event that primes spermatozoa for successful fertilization. Upon induction, acrosomal exocytosis proceeds via a wave of vesiculation that radiates across the sperm head, destabilizing the acrosomal vesicle and resulting in the release of the acrosomal contents. Having shed their acrosome, ...
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Immunology

Metabolomic and Lipidomic Analysis of Bone Marrow Derived Macrophages Authors:  Gretchen L. Seim, Steven V. John and Jing Fan, date: 07/20/2020, view: 889, Q&A: 0
Macrophages are highly plastic immune cells that are capable of adopting a wide array of functional phenotypes in response to environmental stimuli. The changes in macrophage function are often supported and regulated by changes in cellular metabolism. Capturing a comprehensive picture of metabolism is vital for understanding the role of metabolic ...
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Microbiology

Intestinal Enteroid Culture for Human Astroviruses Authors:  Irene A. Owusu, Carmen Mirabelli and Abimbola O. Kolawole, date: 07/20/2020, view: 547, Q&A: 0
Human astroviruses (HAstV) are non-enveloped, positive-sense single stranded RNA viruses that typically cause gastroenteritis in children, the elderly and among immunocompromised individuals. Some HAstV species have also been implicated in neurological diseases. It is important to study these viruses to understand the pathogenesis and develop ...
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A SsrA/NIa-based Strategy for Post-Translational Regulation of Protein Levels in Gram-negative Bacteria Authors:  Gonzalo Durante-Rodríguez, Belén Calles, Víctor de Lorenzo and Pablo I. Nikel, date: 07/20/2020, view: 614, Q&A: 0
Strategies to control the levels of key enzymes of bacterial metabolism are commonly based on the manipulation of gene of interest within the target pathway. The development of new protocols towards the manipulation of biochemical processes is still a major challenge in the field of metabolic engineering. On this background, the FENIX (functional ...
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Neuroscience

Method for Prolonged Incubation of Brain Slices Authors:  Orsolya Kekesi and Yossi Buskila, date: 07/20/2020, view: 581, Q&A: 0
Slices of neuronal tissue maintain a high degree of topographical and functional properties of neurons and glia and therefore are extensively used for measurements of neuronal activity at the molecular, cellular and network levels. However, the lifespan of slice preparations is narrow, averaging of 6-8 hours. Moreover, the average viability of ...
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Calcein Release Assay to Measure Membrane Permeabilization by Recombinant Alpha-Synuclein Authors:  Sayan Dutta, Ben G. Watson, Seema Mattoo and Jean-Christophe Rochet, date: 07/20/2020, view: 800, Q&A: 0
Lipid membranes are involved in regulating biochemical and biological processes and in modulating the selective permeability of cells, organelles, and vesicles. Membrane composition, charge, curvature, and fluidity all have concerted effects on cellular signaling and homeostasis. The ability to prepare artificial lipid assemblies that mimic ...
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Plant Science

RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings Authors:  Tianran Jia and Brandon H. Le, date: 07/20/2020, view: 581, Q&A: 0
Steady-state mRNA levels are determined by both the rates of transcription and degradation. Regulation of mRNA stability and/or degradation are key factors that can significantly affect mRNA levels and its biological functions. mRNA stability can be measured indirectly after transcription inhibition. This protocol described a rapid and sensitive ...
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Gene Expression and Genome Editing Systems by Direct Delivery of Macromolecules Into Rice Egg Cells and Zygotes Authors:  Erika Toda and Takashi Okamoto, date: 07/20/2020, view: 662, Q&A: 0
Polyethylene glycol calcium (PEG-Ca2+)-mediated transfection allows rapid and efficient examination to analyze diverse cellular functions of genes of interest. In plant cells, macromolecules, such as DNA, RNA and protein, are delivered into protoplasts derived from somatic tissues or calli via PEG-Ca2+ transfection. To ...
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Generation and in Planta Functional Analysis of Potato Virus Y mutants Authors:  Katja Stare, Anna Coll, Ion Gutiérrez-Aguirre, Magda Tušek Žnidarič, Maja Ravnikar, Andreja Kežar, Luka Kavčič, Marjetka Podobnik and Kristina Gruden, date: 07/20/2020, view: 570, Q&A: 0
Potato virus Y (PVY), the type member of the genus Potyvirus (family Potyviridae), is the most widespread virus affecting potato and is included in the top five most economically detrimental plant viruses. Recently, the structure of the PVY virion has been determined by cryo-electron microscopy, which has opened the doors to ...
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Systems Biology

HoSeIn: A Workflow for Integrating Various Homology Search Results from Metagenomic and Metatranscriptomic Sequence Datasets Authors:  Gaston Rozadilla, Jorgelina Moreiras Clemente and Christina B. McCarthy, date: 07/20/2020, view: 624, Q&A: 0
Data generated by metagenomic and metatranscriptomic experiments is both enormous and inherently noisy. When using taxonomy-dependent alignment-based methods to classify and label reads, the first step consists in performing homology searches against sequence databases. To obtain the most information from the samples, nucleotide sequences are ...
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