• Volume 11, 2021
  • 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.3890

Past Issues in 2021

Volume: 11 Issue: 2

Biochemistry

A mant-GDP Dissociation Assay to Compare the Guanine Nucleotide Binding Preference of Small GTPases Authors:  Yuping Tan and Qingxiang Sun, date: 01/20/2021, view: 1966, Q&A: 0

Small GTPases are cellular switches that are switched on when bound to GTP and switched off when bound to GDP. Different small GTPase proteins or those with mutations may bind to GTP or GDP with different relative affinities. However, small GTPases generally have very high affinities for guanine nucleotides, rendering it difficult to compare the

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Polyamine Transport Assay Using Reconstituted Yeast Membranes Authors:  Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe, date: 01/20/2021, view: 1359, Q&A: 0

ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure

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Reverse Phase-high-performance Liquid Chromatography (RP-HPLC) Analysis of Globin Chains from Human Erythroid Cells Authors:  Anne Chalumeau, Giacomo Frati, Elisa Magrin and Annarita Miccio, date: 01/20/2021, view: 1123, Q&A: 0

β-hemoglobinopathies are severe genetic disorders characterized either by the abnormal synthesis of the adult β-globin chains of the hemoglobin (Hb) tetramer (βS-globin chains) in sickle cell disease (SCD) or by the reduced β-globin production in β-thalassemia. The identification and quantification of globin chains are

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Cancer Biology

Imaging of Human Cancer Cells in 3D Collagen Matrices Authors:  Karin Pfisterer, Brooke Lumicisi and Maddy Parsons, date: 01/20/2021, view: 2446, Q&A: 0

Research on cell migration and interactions with the extracellular matrix (ECM) was mostly focused on 2D surfaces in the past. Many recent studies have highlighted differences in migratory behaviour of cells on 2D surfaces compared to complex cell migration modes in 3D environments. When embedded in 3D matrices, cells constantly sense the

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Evaluation of Toxicity with Brine Shrimp Assay Authors:  Christina N. Banti and Sotiris K. Hadjikakou, date: 01/20/2021, view: 2337, Q&A: 0

The in vivo toxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) or their hydrogels such as with hydroxyethyl-methacrylate (HEMA) (pHEMA@SBAMs or pHEMA@CoMeDs) are evaluated by the brine shrimp assay. Thus individuals of Artemia salina larvae are incubated in saline solutions with

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Immunology

Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation Reporter-based Detection System Authors:  Laurie G. Landry, Sarah E. Mann, Amanda M. Anderson and Maki Nakayama, date: 01/20/2021, view: 1714, Q&A: 0

Immune tolerance and response are both largely driven by the interactions between the major histocompatibility complex (MHC) expressed by antigen presenting cells (APCs), T-cell receptors (TCRs) on T-cells, and their cognate antigens. Disordered interactions cause the pathogenesis of autoimmune diseases such as type 1 diabetes. Therefore, the

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Wounding Caenorhabditis elegans with Glass Wool Authors:  Murugesan Pooranachithra, James Prabhanand Bhaskar and Krishnaswamy Balamurugan, date: 01/20/2021, view: 1168, Q&A: 0

Research on wound healing majorly relies on rat, mice and other animal models. However, an alternative animal model ought to be brought in the field, pertaining to the stringent ethical issues owing to the use of animals in research. In this regard, Caenorhabdits elegans, a miniature model nematode gains the great attention of the researchers in

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Microbiology

Bacterial Adhesion Kinetics in a High Throughput Setting in Seconds-minutes Time Resolution Authors:  Nimrod Shteindel and Yoram Gerchman, date: 01/20/2021, view: 898, Q&A: 0
Bacterial surface adhesion, the first step in many important processes including biofilm formation and tissue invasion, is a fast process that occurs on a time scale of seconds. Adhesion patterns tend to be stochastic and spatially heterogeneous, especially when bacteria are present in low population densities and at early stages of adhesion to ...
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Determination of Antibacterial Activity of Film Coatings against Four Clinically Relevant Bacterial Strains Authors:  Natalia A. Scilletta, Magdalena Pezzoni, Martín F. Desimone, Galo J. A. A. Soler-Illia, Martín G. Bellino and Paolo N. Catalano, date: 01/20/2021, view: 1410, Q&A: 0

Antibacterial coatings have currently gained great importance in biomedical technology investigations. Because of the spatial arrangement of the film coatings, evaluation of antibacterial activity presents a new challenge regarding traditional bacterial counting methods. In this protocol, four clinically relevant pathogens, Salmonella typhimurium,

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Analysis of Pseudomonas aeruginosa c-di-GMP High and Low Subpopulations Using Flow-assisted Cell Sorting (FACS) and Quantitative Reverse Transcriptase PCR (qRT-PCR) Authors:  Catherine R. Armbruster and Matthew R. Parsek, date: 01/20/2021, view: 1107, Q&A: 0

Cyclic diguanylate monophosphate (c-di-GMP) is a second messenger signaling molecule that drives the transition from planktonic to the biofilm mode of growth in many bacterial species. Pseudomonas aeruginosa has at least two surface sensing systems that produce c-di-GMP in response to surface attachment, the Wsp and Pil-Chp systems. We recently

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Bacterial Conjugation Protocol for Ruminant Mycoplasmas Authors:  Eveline Sagné, Christine Citti and Emilie Dordet-Frisoni, date: 01/20/2021, view: 843, Q&A: 0

In Mycoplasma agalactiae, two simultaneous processes of DNA transfer have been described that require direct cell-to-cell contact and are similar to conjugation. One involves the self-transmission of an integrative conjugative element (ICE) while the second concerns the horizontal transfer of large and small fragments of chromosomal DNA. Here, we

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A One-enzyme RT-qPCR Assay for SARS-CoV-2, and Procedures for Reagent Production Authors:  Sanchita Bhadra, Andre C. Maranhao, Inyup Paik and Andrew D. Ellington, date: 01/20/2021, view: 1075, Q&A: 1

Given the scale of the ongoing COVID-19 pandemic, the need for reliable, scalable testing, and the likelihood of reagent shortages, especially in resource-poor settings, we have developed an RT-qPCR assay that relies on an alternative to conventional viral reverse transcriptases, a thermostable reverse transcriptase/DNA polymerase (RTX) (Ellefson

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Neuroscience

Characterization of Hippocampal Adult-borne Granule Cells in a Transient Cerebral Ischemia Model Authors:  Mihai Ceanga, Madlen Guenther, Ina Ingrisch and Albrecht Kunze, date: 01/20/2021, view: 773, Q&A: 0

Long-term consequences of stroke significantly impair the quality of life in a growing population of stroke survivors. Hippocampal adult neurogenesis has been hypothesized to play a role in the pathophysiology of cognitive and neuropsychiatric long-term sequelae of stroke. Reliable animal models of stroke are paramount to understanding their

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Low- and High-resolution Dynamic Analyses for Magnetic Resonance Spectroscopy Data Author:  Reuben Rideaux, date: 01/20/2021, view: 1335, Q&A: 0

Magnetic resonance spectroscopy (MRS) can be used to measure in vivo concentrations of neurometabolites. This information can be used to identify neurotransmitter involvement in healthy (e.g., perceptual and cognitive processes) and unhealthy brain function (e.g., neurological and psychiatric illnesses). The standard approach for analyzing MRS

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Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex Authors:  Ariel Diaz, Enrique Torre and Manuel Yepes, date: 01/20/2021, view: 1274, Q&A: 1

The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative diseases. Thus, a methodology to study synaptic structure and function is crucial for the

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Plant Science

Cellulase and Macerozyme-PEG-mediated Transformation of Moss Protoplasts Authors:  Rituraj Batth, Ilenuta Simina Cuciurean, Satish Kumar Kodiripaka, Sarah Skovlund Rothman, Caroline Greisen and Henrik Toft Simonsen, date: 01/20/2021, view: 1444, Q&A: 0
This protocol describes the generation of protoplasts from protonemal tissue of the moss Physcomitrium patens (syn. Physcomitrella patens), using Cellulase ONOZUKA R10 and Macerozyme R10, followed by polyethylene glycol (PEG) mediated transformation. The protonemal tissue grown in liquid suspension was harvested and treated with ...
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Design and Construction of Repacked Soil Columns for Measuring Solute Transport, Plant Growth and Soil Biota Authors:  Israel Ikoyi and Achim Schmalenberger, date: 01/20/2021, view: 1071, Q&A: 0

Researchers face a number of challenges in the construction of soil columns which can affect the outcome of their experiments. The use of intact soil cores closely mimics actual field conditions. However, the excavation of large intact soil cores is a time-consuming, labor-intensive process and may lead to soil compaction that would influence the

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Stem Cell

Histological and Immunohistochemical Examination of Stem Cell Proliferation and Reepithelialization in the Wounded Skin Authors:  Rupali Gund, Ravindra Zirmire, Haarshaadri J., Gaurav Kansagara and Colin Jamora, date: 01/20/2021, view: 2362, Q&A: 0

The skin is the largest organ that protects our body from the external environment and it is constantly exposed to pathogenic insults and injury. Repair of damage to this organ is carried out by a complex process involving three overlapping phases of inflammation, proliferation and remodeling. Histological analysis of wounded skin is a convenient

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