• Volume 12, 2022
  • 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.4270

Biochemistry

A Native PAGE Assay for the Biochemical Characterization of G Protein Coupling to GPCRs Authors:  Amanda M. Roehrkasse, Jordan A. Karim and Augen A. Pioszak, date: 12/20/2021, view: 1378, Q&A: 0

G protein-coupled receptors (GPCRs) are a large family of membrane-embedded receptors that have diverse roles in physiology and are major drug targets. GPCRs transduce an agonist binding signal across the membrane to activate intracellular heterotrimeric G proteins. The dynamic nature of the receptors and the complexity of their interactions with

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A Simple and Straightforward Approach for Generating Small, Stable, Homogeneous, Unilamellar 1-Palmitoyl 2-Oleoyl Phosphatidylcholine (POPC) Bilayer Vesicles Authors:  Yong-Guang Gao, Le Thi My Le, Amer Alam and Rhoderick E. Brown, date: 12/20/2021, view: 850, Q&A: 0

Various methods have been developed to generate phosphoglyceride liposomes. Approaches resulting in homogeneous populations of unilamellar bilayer vesicles are generally preferred to mimic various cell membrane situations, as well as to optimize aqueous solute trapping efficiency using the least amount of lipid for biotechnological purposes. Most

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Biophysics

Label-free Quantification of Direct Protein-protein Interactions with Backscattering Interferometry Authors:  Seher Abbas and Karl-Wilhelm Koch, date: 12/20/2021, view: 830, Q&A: 0

The functional performance of a cell depends on how macromolecules, in particular proteins, come together in a precise orientation, how they assemble into protein complexes and interact with each other. In order to study protein-protein interactions at a molecular level, a variety of methods to investigate these binding processes yield affinity

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Copper Based Site-directed Spin Labeling of Proteins for Use in Pulsed and Continuous Wave EPR Spectroscopy Authors:  Kevin Singewald, James A. Wilkinson and Sunil Saxena, date: 12/20/2021, view: 713, Q&A: 0

Site-directed spin labeling in conjunction with electron paramagnetic resonance (EPR) is an attractive approach to measure residue specific dynamics and point-to-point distance distributions in a biomolecule. Here, we focus on the labeling of proteins with a Cu(II)-nitrilotriacetic acid (NTA) complex, by exploiting two strategically placed

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Single Molecular Resolution to Monitor DNA Replication Fork Dynamics upon Stress by DNA Fiber Assay Author:  Wenpeng Liu, date: 12/20/2021, view: 704, Q&A: 0

DNA replication always encounters numerous endogenous and exogenous stresses during the cell cycle. Measuring the cell responses to stress has primarily relied on cell survival and incorporation of radioactive dNTPs, which is limited in resolution. A higher resolution is required to monitor how replication and repair respond to these stresses.

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Cancer Biology

Reconstruction of Human AML Using Functionally and Immunophenotypically Defined Human Haematopoietic Stem and Progenitor Cells as Targeted Populations Authors:  Bernd B. Zeisig, Tsz Kan Fung, Estelle Troadec and Chi Wai Eric So, date: 12/20/2021, view: 1143, Q&A: 0

Acute myeloid leukaemia (AML) is a highly heterogenous blood cancer, in which the expansion of aberrant myeloid blood cells interferes with the generation and function of normal blood cells. Although key driver mutations and their associated inhibitors have been identified in the last decade, they have not been fully translated into better

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Cell Biology

Preparation of Giant Escherichia coli spheroplasts for Electrophysiological Recordings Authors:  Yihao Jiang, Vinay Idikuda and Baron Chanda, date: 12/20/2021, view: 711, Q&A: 0

Prokaryotic ion channels have been instrumental in furthering our understanding of many fundamental aspects of ion channels’ structure and function. However, characterizing the biophysical properties of a prokaryotic ion channel in a native membrane system using patch-clamp electrophysiology is technically challenging. Patch-clamp is

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Developmental Biology

Wounding Zebrafish Larval Epidermis by Laceration Authors:  Andrew S. Kennard, Christopher K. Prinz, Ellen C. Labuz and Julie A. Theriot, date: 12/20/2021, view: 737, Q&A: 0

Wound healing is a critical process for maintaining the integrity of tissues, driven in large part by the active migration of cells to cover damaged regions. While the long-term tissue injury response over hours and days has been extensively studied, the rapid early migratory response of cells to injury in vivo is still being uncovered, especially

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A Method to Induce Brown/Beige Adipocyte Differentiation from Murine Preadipocytes Authors:  Andréa Livia Rocha, Beatriz Alves Guerra, Jeremie Boucher and Marcelo A. Mori, date: 12/20/2021, view: 506, Q&A: 0

Adipocytes exhibit different morphological and functional characteristics, depending on their anatomical location, developmental origin, and stimulus. While white adipocytes tend to accumulate energy as triglycerides, brown and beige adipocytes tend to direct carbon sources to fuel thermogenesis. White and beige adipocytes originate from common

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Measurement of Reactive Oxygen and Nitrogen Species in Living Cells Using the Probe 2',7'-Dichlorodihydrofluorescein Authors:  Waleska Dornas and Detlef Schuppan, date: 12/20/2021, view: 656, Q&A: 0

Reactive oxygen species and reactive nitrogen species (RONS) are involved in programmed cell death in the context of numerous degenerative and chronic diseases. In particular, the ability of cells to maintain redox homeostasis is necessary for an adaptive cellular response to adverse conditions that can cause damage to proteins and DNA, resulting

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Immunology

A Combinatorial in-silico, in-vitro and in-vivo Approach to Quantitatively Study Peptide Induced MHC Stability Authors:  Roman Sarkar, Yashu Sharma, Ayush Jain, Azeez Tehseen, Sudhakar Singh and Sharvan Sehrawat, date: 12/20/2021, view: 464, Q&A: 0

Here, we describe a combinatorial approach in reverse vaccinology to identify immunogenic class I major histocompatibility complex (MHC) displayed epitopes derived from a morbillivirus named pestes des petits ruminants (PPRV). The protocol describes an in silico prediction of immunogenic epitopes using an IEDB tool. The predicted peptides were

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Microbiology

Extraction and Electrophoretic Analysis of Bacterial Lipopolysaccharides and Outer Membrane Proteins Authors:  Yue-Jia Lee and Thomas J. Inzana, date: 12/20/2021, view: 991, Q&A: 0

Lipopolysaccharides (LPS) (or lipooligosaccharides [LOS], which lack the O-antigen side chains characteristic of LPS), and outer membrane proteins (OMP) are major cell-surface molecules in the outer membrane (OM) of gram-negative bacteria. The LPS is responsible for causing endotoxic shock in infected hosts and, in conjunction with some OMPs,

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Selection of Vaccinia Virus Recombinants Using CRISPR/Cas9 Authors:  Anjali Gowripalan, Stewart A. Smith and David C. Tscharke, date: 12/20/2021, view: 505, Q&A: 0

The engineering of poxvirus genomes is fundamental to primary and applied virology research. Indeed, recombinant poxviruses form the basis for many novel vaccines and virotherapies but producing and purifying these viruses can be arduous. In recent years, CRISPR/Cas9 has become the favoured approach for genome manipulation due to its speed and

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Neuroscience

Delivery of AAV for Expression of Fluorescent Biosensors in Juvenile Mouse Hippocampus Authors:  Carlos Manlio Díaz-García, Nidhi Nathwani, Juan Ramón Martínez-François and Gary Yellen, date: 12/20/2021, view: 478, Q&A: 0

Genetically encoded fluorescent biosensors are versatile tools for studying brain metabolism and function in live tissue. The genetic information for these biosensors can be delivered into the brain by stereotaxic injection of engineered adeno-associated viruses (AAVs), which can selectively target different cell types depending on the capsid

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Assessment of Corticospinal Excitability in Awake Rodents Using EMG-Controlled Intracortical Stimulation Authors:  Windsor K. C. Ting, Derek Burns, Maxime Huot-Lavoie and Christian Ethier, date: 12/20/2021, view: 259, Q&A: 0

Assessment of corticospinal excitability (CSE) is an essential component of experiments designed to induce or study neuronal plasticity in the motor system. Common examples are paired associative stimulation (PAS), theta-burst stimulation (TBS), intensive motor training, or any methods aimed at potentiating the corticomotor system in the hope of

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Plant Science

Non-invasive Imaging of Rice Roots in Non-compacted and Compacted Soil Authors:  Bipin K. Pandey, Jonathan A. Atkinson and Craig J. Sturrock, date: 12/20/2021, view: 1212, Q&A: 0

Roots are the prime organ for nutrient and water uptake and are therefore fundamental to the growth and development of plants. However, physical challenges of a heterogeneous environment and diverse edaphic stresses affect root growth in soil. Compacted soil is a serious global problem, causing inhibition of root elongation, which reduces surface

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An in vitro Coupled Assay for PEPC with Control of Bicarbonate Concentration Authors:  Nicholas R. Moody, Chatawal Phansopal and James D. Reid, date: 12/20/2021, view: 352, Q&A: 0

Phosphoenolpyruvate carboxylase (PEPC) catalyzes a critical step in carbon metabolism in plants and bacteria, the irreversible reaction between bicarbonate and phosphoenolpyruvate to produce the C4 compound oxaloacetate. This enzyme is particularly important in the context of C4 photosynthesis, where it is the initial carbon-fixing enzyme. Many

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Bacterial Infection and Hypersensitive Response Assays in Arabidopsis-Pseudomonas syringae Pathosystem Authors:  Minhang Yuan and Xiu-Fang Xin, date: 12/20/2021, view: 587, Q&A: 0

Arabidopsis thaliana-Pseudomonas syringae pathosystem has been used as an important model system for studying plant-microbe interactions, leading to many milestones and breakthroughs in the understanding of plant immune system and pathogenesis mechanisms. Bacterial infection and plant disease assessment are key experiments in the studies of

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