• 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.4209

Biochemistry

Thermal Proteome Profiling to Identify Protein-ligand Interactions in the Apicomplexan Parasite Toxoplasma gondii Authors:  Alice L. Herneisen and Sebastian Lourido, date: 11/05/2021, view: 1359, Q&A: 0

Toxoplasma gondii is a single-celled eukaryotic parasite that chronically infects a quarter of the global population. In recent years, phenotypic screens have identified compounds that block parasite replication. Unraveling the pathways and molecular mechanisms perturbed by such compounds requires target deconvolution. In parasites, such

...
More >>

Biological Engineering

Development of a 3D Bioprinted Scaffold with Spatio-temporally Defined Patterns of BMP-2 and VEGF for the Regeneration of Large Bone Defects Authors:  Fiona E. Freeman, Pierluca Pitacco, Lieke H. A. van Dommelen, Jessica Nulty, David C. Browe, Jung-Youn Shin, Eben Alsberg and Daniel J. Kelly, date: 11/05/2021, view: 978, Q&A: 0

The local delivery of growth factors such as BMP-2 is a well-established strategy for the repair of bone defects. The limitations of such approaches clinically are well documented and can be linked to the need for supraphysiological doses and poor spatio-temporal control of growth factor release in vivo. Using bioprinting techniques, it is

...
More >>

Cancer Biology

Measuring Real-time DNA/RNA Nuclease Activity through Fluorescence Authors:  Paulina Wyrzykowska, Sally Rogers and Richard Chahwan, date: 11/05/2021, view: 1062, Q&A: 0

DNA and RNA nucleases are wide-ranging enzymes, taking part in broad cellular processes from DNA repair to immune response control. Growing interest in the mechanisms and activities of newly discovered nucleases inspired us to share the detailed protocol of our nuclease assay (Sheppard et al., 2019). This easy and inexpensive method can provide

...
More >>

Developmental Biology

Intact in situ Preparation of the Drosophila melanogaster Lymph Gland for a Comprehensive Analysis of Larval Hematopoiesis Authors:  Diana Rodrigues, K. VijayRaghavan, Lucas Waltzer and Maneesha S. Inamdar, date: 11/05/2021, view: 1344, Q&A: 0

Blood cells have a limited lifespan and are replenished by a small number of hematopoietic stem and progenitor cells (HSPCs). Adult vertebrate hematopoiesis occurs in the bone marrow, liver, and spleen, rendering a comprehensive analysis of the entire HSPC pool nearly impossible. The Drosophila blood system is well studied and has developmental,

...
More >>
Targeting the Expression of Long Noncoding RNAs in Murine Satellite Cells from Single Myofibers Authors:  Martina Macino, Beatrice Biferali, Andrea Cipriano, Monica Ballarino and Chiara Mozzetta, date: 11/05/2021, view: 893, Q&A: 0

LncRNAs have been recently implicated in the epigenetic control of muscle differentiation and their functional characterization has traditionally relied upon in vitro models of myogenic differentiation. However, the use of experimental paradigms to specifically target lncRNAs expression in muscle stem cells (MuSCs), also known as satellite cells,

...
More >>
APEX-mediated Proximity Labeling of Proteins in Cells Targeted by Extracellular Vesicles Authors:  Lu Song, Jun Chen, Angela Sun and Randy Schekman, date: 11/05/2021, view: 1030, Q&A: 0

Extracellular vesicles (EVs) are thought to mediate intercellular communication through the delivery of cargo proteins and RNA to target cells. The uptake of EVs is often followed visually using lipophilic-dyes or fluorescently-tagged proteins to label membrane constituents that are then internalized into recipient cells (Christianson et al.,

...
More >>

Immunology

Lentivirus-mediated Conditional Gene Expression Authors:  Leah Rommereim, Ajay Suresh Akhade, Ronald N. Germain, Iain D. C. Fraser and Naeha Subramanian, date: 11/05/2021, view: 970, Q&A: 0

The ability to identify the role of a particular gene within a system is dependent on control of the expression of that gene. In this protocol, we describe a method for stable, conditional expression of Nod-Like receptors (NLRs) in THP-1 cells using a lentiviral expression system. This system combines all the necessary components for

...
More >>
Suppression of Human Dendritic Cells by Regulatory T Cells Authors:  Qing Huang, Avery J. Lam, Dominic A. Boardman, Nicholas A. J. Dawson and Megan K. Levings, date: 11/05/2021, view: 771, Q&A: 0

Regulatory T cells (Tregs) suppress immune responses via a variety of mechanisms and can be used as a cellular therapy to induce tolerance. The function of Tregs is commonly assessed in vitro using assays that measure suppression of effector T cell proliferation and/or cytokine production. However, Tregs can also suppress the function of antigen

...
More >>
Production of Recombinant Replication-defective Lentiviruses Bearing the SARS-CoV or SARS-CoV-2 Attachment Spike Glycoprotein and Their Application in Receptor Tropism and Neutralisation Assays Authors:  Nazia Thakur, Giulia Gallo, Ahmed M. E. Elreafey and Dalan Bailey, date: 11/05/2021, view: 706, Q&A: 0

For enveloped viruses, such as SARS-CoV-2, transmission relies on the binding of viral glycoproteins to cellular receptors. Conventionally, this process is recapitulated in the lab by infection of cells with isolated live virus. However, such studies can be restricted due to the availability of high quantities of replication-competent virus,

...
More >>

Microbiology

An Assay to Determine NAD(P)H: Quinone Oxidoreductase Activity in Cell Extracts from Candida glabrata Authors:  Anamika Battu, Rajaram Purushotham and Rupinder Kaur, date: 11/05/2021, view: 503, Q&A: 0

Flavodoxin-like proteins (Fld-LPs) are an important constituent of the oxidative stress defense system in several organisms and highly conserved from bacteria to humans. These proteins possess NAD(P)H:quinone oxidoreductase activity and convert quinones to hydroquinones through two-electron reduction, using NAD(P)H and quinone as electron donor

...
More >>
Quantification of Duloxetine in the Bacterial Culture and Medium to Study Drug-gut Microbiome Interactions Authors:  Prasad B. Phapale, Sonja Blasche, Kiran R. Patil and Theodore Alexandrov, date: 11/05/2021, view: 515, Q&A: 0

Expanding our understanding of drug-gut bacteria interactions requires high-throughput drug measurements in complex bacterial cultures. Quantification of drugs in the cultures, media, and cell pellets is prone to strong matrix effects. We have developed a liquid chromatography–high resolution mass spectrometry (LC–HRMS) method for

...
More >>
Determination of Fungal Tolerance Index to Heavy Metals and Heavy Metal Resistance Tests Author:  Rosa Paulina Calvillo-Medina, date: 11/05/2021, view: 472, Q&A: 0

Fungal metallo-tolerance has been described in different species and plays an important role in bioremediation of contaminated environments. Metallo-tolerance is mainly documented by microdilution assays and agar well diffusion methods using equipment that can be expensive. The tolerance index can be calculated to determine the efficiency of a

...
More >>

Molecular Biology

Enrichment of Cytoplasmic RNA Granules from Arabidopsis Seedlings Authors:  Zhen Lei, Eun Yu Kim and Jungnam Cho, date: 11/05/2021, view: 576, Q&A: 0

RNA granules (RGs) are membraneless intracellular compartments that play important roles in the post-transcriptional control of gene expression. Stress granules (SGs) are a type of RGs that form under environmental challenges and/or internal cellular stresses. Stress treatments lead to strong mRNAs translational inhibition and storage in SGs until

...
More >>
Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes Authors:  Cecilia Di Genova, Alex Sampson, Simon Scott, Diego Cantoni, Martin Mayora-Neto, Emma Bentley, Giada Mattiuzzo, Edward Wright, Mariliza Derveni, Bethany Auld, Bill T. Ferrara, Dale Harrison, Mohamed Said, Arwa Selim, Elinor Thompson, Craig Thompson, George Carnell and Nigel Temperton, date: 11/05/2021, view: 794, Q&A: 0

This protocol details a rapid and reliable method for the production and titration of high-titre viral pseudotype particles with the SARS-CoV-2 spike protein (and D614G or other variants of concern, VOC) on a lentiviral vector core, and use for neutralisation assays in target cells expressing angiotensin-converting enzyme 2 (ACE2) and

...
More >>
A Nucleocapsid-based Transcomplementation Cell Culture System of SARS-CoV-2 to Recapitulate the Complete Viral Life Cycle Authors:  Yanying Yu, Xiaohui Ju and Qiang Ding, date: 11/05/2021, view: 488, Q&A: 0

The ongoing COVID-19 pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As this virus is classified as a biosafety level-3 (BSL-3) agent, the development of countermeasures and basic research methods is logistically difficult. Recently, using reverse genetics, we developed a BSL-2 cell culture system for production

...
More >>

Neuroscience

Whole-mount Staining of Mouse Diaphragm Neuromuscular Junctions Authors:  Rula Sha, Zhenzhen Wang, Xue You, Yuguo Liu, Zhiqin Xie and Ying Feng, date: 11/05/2021, view: 445, Q&A: 0

The neuromuscular junction (NMJ) is a specialized synapse that connects the terminal end of a motor neuron and a skeletal muscle fiber. Defects in NMJ cause abnormalities of neuromuscular transmission, leading to NMJ disorders. The mammalian diaphragm muscle is essential for respiration and has been widely used to study NMJ formation. Here, we

...
More >>

Plant Science

Preference Test of Plutella xylostella Larvae upon DMNT Treatment Authors:  Chen Chen, Zhen Tao, Chuanhong Wang, Tengyue Wang, Hongyi Chen, Taoshan Jiang and Peijin Li, date: 11/05/2021, view: 417, Q&A: 0

We describe a method to test the preference of insects in response to (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). We use a device that includes a horizontal glass tube, two grooves (with activated carbon), air flow, rubber stoppers/tubes, transparent glass containers (optional), and a holder for the glass tube (optional). Equal amounts of activated

...
More >>

Stem Cell

High-throughput 3D Spheroid Formation and Effective Cardiomyocyte Differentiation from Human iPS Cells Using the Microfabric Vessels EZSPHERETM Authors:  Tatsuaki Miwa, Alimjan Idiris and Hiromichi Kumagai, date: 11/05/2021, view: 555, Q&A: 0

High-throughput 3D spheroid formation from human induced pluripotent stem cells (hiPSCs) can be easily performed using the unique microfabric vessels EZSPHERE, resulting in effective and large scale generation of differentiated cells such as cardiomyocytes or neurons. Such hiPSC-derived cardiomyocytes (hiPSC-CMs) or neurons are very useful in the

...
More >>

Systems Biology

Histone modification ChIP-seq on Arabidopsis thaliana Plantlets Authors:  Juan S. Ramirez-Prado, David Latrasse and Moussa Benhamed, date: 11/05/2021, view: 631, Q&A: 1

Characterizing the molecular mechanisms regulating gene expression is crucial for understanding the regulatory processes underlying physiological responses to environmental and developmental signals in eukaryotes. The covalent modification of histones contributes to the compaction levels of chromatin, as well as the recruitment of the

...
More >>
We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.