• 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.3594

Past Issues in 2020

Volume: 10 Issue: 8

Biochemistry

Fibrin Breakdown Assay Authors:  Jiayue Ling, Connor M. Blair and George S. Baillie, date: 04/20/2020, view: 1282, Q&A: 0
Fibrinolysis is an integral part of the matrix remodeling process that contributes to tissue repair. Fibrin clots are broken down during fibrinolysis in a controlled process. Fibrin degradation products (FDPs) have also been shown to have a role in the regulation of cell growth and are implicated in various vascular diseases. This protocol was ...
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Enzymatic Construction of Protein Polymer/Polyprotein Using OaAEP1 and TEV Protease Authors:  Yibing Deng, Shengchao Shi, Bin Zheng, Tao Wu and Peng Zheng, date: 04/20/2020, view: 1304, Q&A: 0
The development of chemical and biological coupling technologies in recent years has made possible of protein polymers engineering. We have developed an enzymatic method for building polyproteins using a protein ligase OaAEP1 (asparagine endopeptidase 1) and protease TEV (tobacco etching virus). Using a mobile TEV protease site compatible with the ...
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Cancer Biology

Single Cell Migration Assay Using Human Breast Cancer MDA-MB-231 Cell Line Authors:  David M. Gau and Partha Roy, date: 04/20/2020, view: 1594, Q&A: 0
Cell migration is a fundamental cellular process that plays a crucial role in many physioglogical and pathological processes such as wound healing or cancer metastasis. Many assays have been developed to examine cell migration, such as the wound healing or scratch assay, Boyden Chamber or transwell assay, and the method we will describe here, ...
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Cell Biology

Dual Fluorescence Cytometry Assay to Assess Cellular Protein Levels Authors:  Ashley M Ngo, Ruofan Wang and Andreas S Puschnik, date: 04/20/2020, view: 1244, Q&A: 0
Expression levels of cellular proteins can be affected by various perturbations, such as genetic knockout of interactors, drug treatments or cell stress. To specifically measure the effects on protein levels post-synthesis under different experimental conditions, it is important to compensate for transcriptional and other upstream changes. Here, ...
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Developmental Biology

Auxin-mediated Protein Degradation in Caenorhabditis elegans Authors:  Michael A. Q. Martinez and David Q. Matus, date: 04/20/2020, view: 1348, Q&A: 0
The auxin-inducible degron (AID) technology was recently adapted for use in the nematode Caenorhabditis elegans. Rapid degradation of C. elegans proteins tagged with an AID is mediated by a plant-specific F-box protein, transport inhibitor response 1 (TIR1), and occurs only in the presence of the phytohormone auxin. The first ...
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Immunology

A Triple-challenge Mouse Model of Allergic Airway Disease, Primary Influenza Infection, and Secondary Bacterial Infection Authors:  Sean Roberts, Clare M Williams, Sreeja Roy and Yoichi Furuya, date: 04/20/2020, view: 705, Q&A: 0
Asthma is a global problem that affects millions of individuals. An increased risk of respiratory viral and bacterial infections is one of the complications of asthma. We recently reported that mice with ovalbumin-induced allergic airway disease (AAD) are protected against influenza-Streptococcus pneumoniae co-infection. Here, we describe ...
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Environmental Conditioning and Aerosol Infection of Mice Authors:  Eriko Kudo and Akiko Iwasaki, date: 04/20/2020, view: 1015, Q&A: 0
Influenza infection models in mice are widely used to study flu-mediated immune responses and pathology. However, most laboratory mice are housed at 20 °C and 50% relative humidity (RH). To better recapitulate influenza epidemics and immune responses during winter seasons, mice were housed at 20 °C under different humidity conditions, 10-20% or ...
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Microbiology

In vivo Quantification of Alkanes in Escherichia coli Authors:  Tabinda Shakeel, Zia Fatma and Syed Shams Yazdani, date: 04/20/2020, view: 1106, Q&A: 0
Microbial production of alkanes employing synthetic biology tools has gained tremendous attention owing to the high energy density and similarity of alkanes to existing petroleum fuels. One of the most commonly studied pathways includes the production of alkanes by AAR (acyl-ACP (acyl carrier protein) reductase)-ADO (aldehyde deformylating ...
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Molecular Biology

Isolation and Characterization of Exosomes from Mouse Feces Authors:  Chunhua Yang, Mingzhen Zhang, Junsik Sung, Lixin Wang, Yunjin Jung and Didier Merlin, date: 04/20/2020, view: 1236, Q&A: 0
Exosomes secreted by colonic epithelial cells are present in feces and contain valuable epigenetic information, such as miRNAs, proteins, and metabolites. An in-depth study of this information is conducive to the diagnosis or treatment of relevant diseases. A crucial prerequisite of such a study is to establish an efficient isolation method, ...
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An in vitro Assay to Screen for Substrates of PKA Using Phospho-motif-specific Antibodies Authors:  David M. Gau and Partha Roy, date: 04/20/2020, view: 833, Q&A: 0
Kinases function as regulators of many cellular processes such as cell migration. These enzymes typically phosphorylate target motif sequences. Mass spec or phospho-specific antibody detection can be used to determine whether a kinase can phosphorylate proteins of interest, however, mass spec can be expensive and phospho-antibodies for the protein ...
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High-throughput Flow Cytometry Assay to Investigate TDP43 Splicing Function Authors:  H. Broder Schmidt and Rajat Rohatgi, date: 04/20/2020, view: 1203, Q&A: 0
Mutations in RNA-binding proteins (RBPs) such as TDP43 are associated with transcriptome-wide splicing defects and cause severe neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The impact of RBP mutations on splicing function is routinely studied using PCR-based bulk measurements. ...
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Neuroscience

Minimally Invasive Oral Surgery Induction of the FRICT-ION Chronic Neuropathic Pain Model Authors:  Marena A. Montera and Karin N. Westlund, date: 04/20/2020, view: 821, Q&A: 0
An easily induced preclinical trigeminal neuropathic nerve injury model is described here for the study of chronic pain, the model acronym FRICT-ION (Foramen Rotundum Inflammatory Constriction Trigeminal InfraOrbital Nerve). In patients, neuropathic pain is thought to be related to ...
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Examining Cocaine Conditioning Place Preference in Mice Authors:  MaryElizabeth J. Simkevich, Rianne R. Campbell and André O. White, date: 04/20/2020, view: 726, Q&A: 0
A key component of combating substance use disorders is understanding the neural mechanisms that support drug reward. Tasks such as self-administration assess the reinforcing properties of a drug using a learned behavior but require numerous training sessions and surgery. In comparison, the conditioned place preference (CPP) task assesses reward ...
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Plant Science

MAMP-triggered Medium Alkalinization of Plant Cell Cultures Authors:  Gabriel L. Fiorin, Andrea Sánchez-Vallet, Bart P. H. J. Thomma, Gonçalo A. G. Pereira and Paulo J.P.L. Teixeira, date: 04/20/2020, view: 1020, Q&A: 0
Plants recognize a wide variety of microbial molecules to detect and respond to potential invaders. Recognition of Microbe-Associated Molecular Patterns (MAMPs) by cell surface receptors initiate a cascade of biochemical responses that include, among others, ion fluxes across the plasma membrane. A consequence of such event is a decrease in the ...
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Systems Biology

Circadian Gene Profiling in Laser Capture Microdissected Mouse Club Cells Authors:  Zhenguang Zhang and Andrew Loudon, date: 04/20/2020, view: 880, Q&A: 0
Cell heterogeneity is high in tissues like lung. Research conducted on pure population of cells usually offers more insights than bulk tissues, such as circadian clock work. In this protocol, we provide a detailed work flow on how to do circadian clock study by RNA seq in laser capture micro-dissected mouse lung club cells. The method uses frozen ...
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