• 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.3873

Current Issue in 2021

Volume: 11 Issue: 1

Cancer Biology

A Rigorous Quantitative Approach to Analyzing Phagocytosis Assays Authors:  Michael D. Caponegro, Kaitlyn Koenig Thompson, Maryam Tayyab and Stella E. Tsirka, date: 01/05/2020, view: 1302, Q&A: 0
Studying monocytic cells in isolated systems in vitro contributes significantly to the understanding of innate immune physiology. Functional assays produce read outs which can be used to measure responses to selected stimuli, such as pathogen exposure, antigen loading, and cytokine stimulation. Integration of these results with high ...
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A Transient Transfection-based Cell Adhesion Assay with 293T Cells Authors:  Rohit Singh and Beom K. Choi, date: 01/05/2021, view: 488, Q&A: 0

The in vitro cell adhesion assay is a quantitative method for measuring selective cell adhesion to specific proteins. Traditionally, cell adhesion assays employ purified protein immobilized on a solid glass or plastic surface. Here, we describe a transient 293T cell transfection-based cell adhesion assay to study selective cell adhesion of a

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CENP-C Phosphorylation by CDK1 in vitro Authors:  Reito Watanabe, Masatoshi Hara, Mariko Ariyoshi and Tatsuo Fukagawa, date: 01/05/2021, view: 346, Q&A: 0

Accurate chromosome segregation during mitosis requires the kinetochore, a large protein complex, which makes a linkage between chromosomes and spindle microtubes. An essential kinetochore component, CENP-C, is phosphorylated by Cyclin-B-Cyclin dependent kinase 1 (CDK1) that is a master kinase for mitotic progression, promoting proper kinetochore

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Cell Biology

A Simple Microplate Assay for Reactive Oxygen Species Generation and Rapid Cellular Protein Normalization Authors:  Neville S. Ng and Lezanne Ooi, date: 01/05/2021, view: 672, Q&A: 0
2',7'-dichlorofluorescein (DCF) and derivatives are commonly used as fluorescent indicators of a broad spectrum of reactive oxygen species (ROS) generation in cell-based assays. However, there are numerous challenges inherent to the utilization of DCF probes for intracellular microscopic analysis, including bleaching and probe efflux. Plate ...
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A Parkinson’s Disease-relevant Mitochondrial and Neuronal Morphology High-throughput Screening Assay in LUHMES Cells Authors:  Tom Leah, Irina Vazquez-Villaseñor, Laura Ferraiuolo, Stephen B Wharton and Heather Mortiboys, date: 01/05/2021, view: 425, Q&A: 0

Parkinson’s disease is a devastating neurodegenerative disorder affecting 2-3% of the population over 65 years of age. There is currently no disease-modifying treatment. One of the predominant pathological features of Parkinson’s disease is mitochondrial dysfunction, and much work has aimed to identify therapeutic compounds which can

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Developmental Biology

Measuring Bone Volume at Multiple Densities by Micro-computed Tomography Authors:  Emma C. Walker, Narelle E. McGregor, Audrey S. M. Chan and Natalie A. Sims, date: 01/05/2021, view: 595, Q&A: 0

Bone strength is controlled by both bone mass, and the organization and quality of the bone material. The current standard method for measuring bone mass in mouse and rat studies is micro-computed tomography. This method typically uses a single threshold to identify bone material in the cortical and trabecular regions. However, this single

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Microbiology

Assessment of Diadenylate Cyclase and c-di-AMP-phosphodiesterase Activities Using Thin-layer and Ion Exchange Chromatography Authors:  Andreas Latoscha, David Jan Drexler, Gregor Witte and Natalia Tschowri, date: 01/05/2021, view: 346, Q&A: 0

All living cells use cyclic nucleotides as second messengers for signal sensing and transduction. Cyclic di-3′,5′-adenosine monophosphate (c-di-AMP) is primarily involved in the control of bacterial and euryarcheal osmoadaptation and is produced by diadenylate cyclases from two molecules of ATP. Specific phosphodiesterases hydrolyze

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Neuroscience

Calcium Imaging of Neuronal Activity under Gradually Changing Odor Stimulation in Caenorhabditis elegans Authors:  Yuki Tanimoto and Koutarou D. Kimura, date: 01/05/2021, view: 320, Q&A: 0

Olfactory behavior is among the most fundamental animal behaviors both in the wild and in the laboratory. To elucidate the neural mechanisms underlying olfactory behavior, it is critical to measure neural responses to odorant concentration changes resembling those that animals actually sense during olfactory behavior. However, reproducing the

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Optimized Immunostaining of Embryonic and Early Postnatal Mouse Brain Sections Authors:  Kawssar Harb, Michele Bertacchi and Michèle Studer, date: 01/05/2021, view: 565, Q&A: 0

The mammalian neocortex, the outer layer of the cerebrum and most recently evolved brain region, is characterized by its unique areal and laminar organization. Distinct cortical layers and areas can be identified by the protein expression of graded transcription factors and molecular determinants that define the identity of different projection

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Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis Authors:  Steffen Mayerl and Charles ffrench-Constant, date: 01/05/2021, view: 370, Q&A: 0

The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain

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Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer Authors:  Sung-Jin Lee and Soo-Young Kim, date: 01/05/2021, view: 232, Q&A: 0

The subretinal layer between retinal pigment epithelium (RPE) and photoreceptors is a region involved in inflammation and angiogenesis during the procession of diseases such as age-related macular degeneration. The current protocols of whole mounts (retina and RPE) are unable to address the intact view of the subretinal layer because the

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Plant Science

Non-radioactive Assay to Determine Product Profile of Short-chain Isoprenyl Diphosphate Synthases Authors:  Avanish Rai and Dinesh A. Nagegowda, date: 01/05/2021, view: 332, Q&A: 0
Isoprenoids represent the largest class of metabolites with amazing diversities in structure and function. They are involved in protecting plants against pathogens or herbivores or involved in attracting pollinators. Isoprenoids are derived from geranyl diphosphate (GPP; C10), farnesyl diphosphate (FPP; C15), geranylgeranyl ...
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Development and Standardization of Rapid and Efficient Seed Germination Protocol for Cannabis sativa Authors:  Aleksei Sorokin, Narendra Singh Yadav, Daniel Gaudet and Igor Kovalchuk, date: 01/05/2021, view: 452, Q&A: 0

Cannabis seed germination is an important process for growers and researchers alike. Many biotechnological applications require a reliable sterile method for seed germination. This protocol outlines a seed germination procedure for Cannabis sativa using a hydrogen peroxide (H2O2) solution as liquid germination media. In this protocol, all three

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Stable Transformation of Arabidopsis thaliana Cell Suspension Cultures: A Case Study for the Overexpression of The COI1 Receptor Authors:  Eva K. Díaz-Sánchez, José A. O’Brien, Imma Pérez-Salamó, Jovaras Krasauskas, Moritz Bömer and Alessandra Devoto, date: 01/05/2021, view: 603, Q&A: 0
Cell suspension cultures have been studied for decades to produce natural molecules. However, the difficulty in generating stably transformed cell lines has limited their use to produce high value chemicals reproducibly and in elevated quantities.

In this protocol, a method to stably transform and maintain Arabidopsis cell ...
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A Pulse–chase EdU Method for Detection of Cell Division Orientation in Arabidopsis and Juncus prismatocarpus Leaf Primordia Authors:  Xiaofeng Yin and Hirokazu Tsukaya, date: 01/05/2021, view: 262, Q&A: 0
In plants, the morphological diversity of leaves is largely determined by cell division, especially cell division orientation. Whereas cell division itself is easily monitored, the detection and quantification of cell division orientation are difficult. The few existing methods for detection and quantification of cell division orientation are ...
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Efficient Transient Gene Knock-down in Tobacco Plants Using Carbon Nanocarriers Authors:  Gozde S. Demirer and Markita P. Landry, date: 01/05/2021, view: 359, Q&A: 0
Gene knock-down in plants is a useful approach to study genotype-phenotype relationships, render disease resistance to crops, and enable efficient biosynthesis of molecules in plants. Small interfering RNA (siRNA)-mediated gene silencing is one of the most common ways to achieve gene knock-down in plants. Traditionally, siRNA is delivered into ...
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