• 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.3846

Biochemistry

Triacylglycerol Measurement in HeLa Cells Authors:  Ximing Du and Hongyuan Yang, date: 12/20/2020, view: 1040, Q&A: 0

Lipid droplets store triacylglycerols (triglycerides) and sterol esters to regulate lipid and energy homeostasis. Triacylglycerol measurement is often performed during the investigation of lipid droplet formation and growth. This protocol describes a reliable method using a fluorometric lipid quantification kit to measure triacylglycerols

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Preparation of Bacterial Outer Membrane Vesicles for Characterisation of Periplasmic Proteins in Their Native Environment Authors:  Johannes Thoma and Björn M. Burmann, date: 12/20/2020, view: 1103, Q&A: 0

Bacterial outer membrane vesicles (OMVs) are naturally formed by budding from the outer membrane of Gram-negative bacteria. OMVs consist of a lipid bilayer identical in composition to the original outer membrane and contain periplasmic content within their lumen. Enriched with specific envelope proteins, OMVs make for an excellent native-like

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Quantitative Irreversible Tethering (qIT) for Target-directed Covalent Fragment Screening Authors:  Gregory B. Craven, David J. Mann and Alan Armstrong, date: 12/20/2020, view: 624, Q&A: 0

Small molecules that react to form covalent bonds with proteins are widely used as biological tools and therapeutic agents. Screening cysteine-reactive fragments against a protein target is an efficient way to identify chemical starting points for covalent probe development. Mass spectrometry is often used to identify the site and degree of

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Cancer Biology

Equilibrium and Kinetic Measurements of Ligand Binding to HiBiT-tagged GPCRs on the Surface of Living Cells Authors:  Michelle E. Boursier, Sergiy Levin, Robin Hurst and Rachel Friedman Ohana, date: 12/20/2020, view: 1024, Q&A: 0

G-protein coupled receptors (GPCRs) remain at the forefront of drug discovery efforts. Detailed assessment of features contributing to GPCR ligand engagement in a physiologically relevant environment is imperative to the development of new therapeutics with improved efficacy. Traditionally, binding properties such as affinity and kinetics were

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Developmental Biology

Whole-mount Immunohistochemistry of Adult Zebrafish Retina for Advanced Imaging Authors:  Mikiko Nagashima and Peter F. Hitchcock, date: 12/20/2020, view: 1047, Q&A: 0

Immunohistochemistry is a widely used technique to examine the expression and subcellular localization of proteins. This technique relies on the specificity of antibodies and requires adequate penetration of antibodies into tissues. The latter is especially challenging for thick specimens, such as embryos and other whole-mount preparations. Here

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Immunology

RNA ImmunoGenic Assay: A Method to Detect Immunogenicity of in vitro Transcribed mRNA in Human Whole Blood Authors:  AKM Ashiqul Haque, Petra Weinmann, Sumit Biswas, Rupert Handgretinger, Markus Mezger, Michael S. D. Kormann and Justin S Antony, date: 12/20/2020, view: 1199, Q&A: 0
The mRNA therapeutics is a new class of medicine to treat many various diseases. However, in vitro transcribed (IVT) mRNA triggers immune responses due to recognition by human endosomal and cytoplasmic RNA sensors, but incorporation of modified nucleosides have been shown to reduce such responses. Therefore, an assay signifying important ...
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Pea Aphid Rearing, Bacterial Infection and Hemocyte Phagocytosis Assay Authors:  Li Ma, Lu Liu and Zhiqiang Lu, date: 12/20/2020, view: 719, Q&A: 0

Insects rely on the simple but effective innate immune system to combat infection. Cellular and humoral responses are interconnected and synergistic in insects’ innate immune system. Phagocytosis is one major cellular response. It is difficult to collect clean hemolymph from the small insect like pea aphid. Here, we provide a practicable

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Microbiology

Preparation of Nippostrongylus brasiliensis Larvae for the Study of Host Skin Response Authors:  Tiffany Bouchery, Gillian Coakley and Nicola L. Harris, date: 12/20/2020, view: 610, Q&A: 0

Hookworms are skin penetrating parasites, however in the laboratory the hookworm model Nippostrongylus brasiliensis, the parasite is traditionally administered subcutaneously bypassing the skin (epidermis and dermis). Here, we describe two complementary approaches for infecting mice with N. brasiliensis in order to study the skin immune responses.

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Antimicrobial Sensitivity Assay for Bdellovibrio bacteriovorus Authors:  Emanuele Marine and Klaas M. Pos, date: 12/20/2020, view: 819, Q&A: 0

Bdellovibrio bacteriovorus, an obligate predatory bacterium [i.e., bacteria that kill and feed on other bacteria (prey)], has the potential to be used as a probiotic for the disinfection of surfaces or for the treatment of bacterial infections. One option is to use this organism in combination with antimicrobials to potentiate the effectiveness of

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Evaluation of the Sequence Variability within the PCR Primer/Probe Target Regions of the SARS-CoV-2 Genome Authors:  Kashif Aziz Khan and Peter Cheung, date: 12/20/2020, view: 1293, Q&A: 0
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; initially named 2019-nCoV) is responsible for the recent coronavirus disease (COVID-19) pandemic, and polymerase chain reaction (PCR) is the current standard method for diagnosis from patient samples. As PCR assays are prone to sequence mismatches due to mutations in the viral genome, it ...
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Neuroscience

A Quantitative Assay to Measure Stress Granule Association of Proteins and Peptides in Semi-permeabilized Human Cells Authors:  Saskia Hutten and Dorothee Dormann, date: 12/20/2020, view: 1245, Q&A: 0

Stress granules (SGs) are membrane-less organelles that form in the cytoplasm through phase separation, in response to diverse stressors. SGs contain translationally stalled mRNAs, proteins involved in translation, and various RNA-binding proteins (RBPs). Due to the high local concentration of aggregation-prone RBPs, SGs might act as condensation

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Nestlet Shredding and Nest Building Tests to Assess Features of Psychiatric Disorders in Mice Authors:  Fabian Dorninger, Gerhard Zeitler and Johannes Berger, date: 12/20/2020, view: 789, Q&A: 0

Mimicking the various facets of human psychiatric and neurodevelopmental disorders in animal models is a challenging task. Nevertheless, mice have emerged as a widely used model system to study pathophysiology and treatment strategies for these diseases. However, the corresponding behavioral tests are often elaborate and require extensive

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Plant Science

Analysis of Isotopically-labeled Monogalactosyldiacylglycerol Molecular Species from [14C]Acetate-Labeled Tobacco Leaves Authors:  Hari Kiran Kotapati and Philip D. Bates, date: 12/20/2020, view: 659, Q&A: 0

Plant lipid metabolism is a dynamic network where synthesis of essential membrane lipids overlaps with synthesis of valuable storage lipids (e.g., vegetable oils). Monogalactosyldiacylglycerol (MGDG) is a key component of the chloroplast membrane system required for photosynthesis and is produced by multiple pathways within the lipid metabolic

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Lipid Droplet Isolation from Arabidopsis thaliana Leaves Authors:  Yovanny Izquierdo, Rubén Fernández-Santos, Tomás Cascón and Carmen Castresana, date: 12/20/2020, view: 938, Q&A: 0

Lipid droplets (LDs) are neutral lipid aggregates surrounded by a phospholipid monolayer and specific proteins. In plants, they play a key role as energy source after seed germination, but are also formed in vegetative tissues in response to developmental or environmental conditions, where their functions are poorly understood. To elucidate these,

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Stem Cell

Differentiation of Human Induced Pluripotent Stem Cells (hiPSCs) into Osteoclasts Author:  I-Ping Chen, date: 12/20/2020, view: 819, Q&A: 0

Defects in bone resorption by osteoclasts result in numerous rare genetic bone disorders as well as in some common diseases such as osteoporosis or osteopetrosis. The use of hiPSC-differentiated osteoclasts opens new avenues in this research field by providing an unlimited cell source and overcoming obstacles such as unavailability of human

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Isolation of Extracellular Vesicles Derived from Mesenchymal Stromal Cells by Ultracentrifugation Authors:  María José Ramírez-Bajo, Elisenda Banon-Maneus, Jordi Rovira, Josep M. Campistol and Fritz Diekmann, date: 12/20/2020, view: 944, Q&A: 0
Extracellular vesicles (EVs) are a heterogeneous group of membranous vesicles that differ on their biogenesis and release pathways, such as exosomes, microvesicles and apoptotic bodies. They are involved in cell-to-cell communication delivering signal molecules (proteins, nucleic acids, lipids, etc.) that can regulate different ...
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