• 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.4441

Past Issues in 2022

Volume: 12 Issue: 11

Biochemistry

H2O2 Release Assay Authors:  Santosh Yadav, Shruthi Sanjitha Sampath, Brian J. Deskin and Victor J. Thannickal, date: 06/05/2022, view: 1097, Q&A: 0

Reactive oxygen species are ubiquitous in nature, and function as signalling molecules in biological systems; they may also contribute to oxidative stress in several pathobiological disease states. In this report, we describe a simple, reliable, sensitive, and specific assay for the detection and quantitation of hydrogen peroxide (H2O2) release by

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Biological Engineering

Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering Authors:  Zhongwen Chen, Kabir H. Biswas and Jay T. Groves, date: 06/05/2022, view: 769, Q&A: 0

A multitude of membrane-localized receptors are utilized by cells to integrate both biochemical and physical signals from their microenvironment. The clustering of membrane receptors is widely presumed to have functional consequences for subsequent signal transduction. However, it is experimentally challenging to selectively manipulate receptor

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Cancer Biology

Plasma Membrane Wounding and Repair Assays for Eukaryotic Cells Authors:  Stine Lauritzen Sønder, Malene Laage Ebstrup, Catarina Dias, Anne Sofie Busk Heitmann and Jesper Nylandsted, date: 06/05/2022, view: 717, Q&A: 0

Damage to the plasma membrane and loss of membrane integrity are detrimental to eukaryotic cells. It is, therefore, essential that cells possess an efficient membrane repair system to survive. However, the different cellular and molecular mechanisms behind plasma membrane repair have not been fully elucidated. Here, we present three complementary

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Cell Biology

Labelling of Active Transcription Sites with Argonaute NRDE-3—Image Active Transcription Sites in vivo in Caenorhabditis elegans Authors:  Antoine Barrière and Vincent Bertrand, date: 06/05/2022, view: 509, Q&A: 0

Live labelling of active transcription sites is critical to our understanding of transcriptional dynamics. In the most widely used method, RNA sequence MS2 repeats are added to the transcript of interest, on which fluorescently tagged Major Coat Protein binds, and labels transcription sites and transcripts. Here we describe another strategy, using

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Electroporation of Small Interfering RNAs into Tibialis Anterior Muscles of Mice Authors:  Anna Stephan, Flavia A. Graca, Liam C. Hunt and Fabio Demontis, date: 06/05/2022, view: 780, Q&A: 0

Aging and wasting of skeletal muscle reduce organismal fitness. Regrettably, only limited interventions are currently available to address this unmet medical need. Many methods have been developed to study this condition, including the intramuscular electroporation of DNA plasmids. However, this technique requires surgery and high electrical

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Developmental Biology

Simple Methods for Permanent or Transient Denervation in Mouse Sciatic Nerve Injury Models Authors:  Alexis Osseni, Jean-Luc Thomas, Alireza Ghasemizadeh, Laurent Schaeffer and Vincent Gache, date: 06/05/2022, view: 594, Q&A: 0

Our ability to move and breathe requires an efficient communication between nerve and muscle that mainly takes place at the neuromuscular junctions (NMJs), a highly specialized synapse that links the axon of a motor neuron to a muscle fiber. When NMJs or axons are disrupted, the control of muscle fiber contraction is lost and muscle are paralyzed.

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Efficient Superovulation and Egg Collection from Mice Authors:  Miyuki Shindo, Kenji Miyado, Woojin Kang, Maki Fukami and Mami Miyado, date: 06/05/2022, view: 547, Q&A: 0

Superovulation is a method used to reduce the number of mice used per experiment by increasing the egg number. Conventionally, superovulation for obtaining mouse eggs involves the use of equine chorionic gonadotropin (eCG) for stimulation and human CG for induction. Female mice of the C57BL/6 inbred strain spontaneously ovulate approximately 10

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Click-iT® Plus OPP Alexa Fluor® Protein Synthesis Assay in Embryonic Cells Authors:  Yan Li, Xu Ji, Lu Chang, Jianan Tang, Min-Min Hua, Jing Liu, Christopher O’Neil, Xuefeng Huang and Xingliang Jin, date: 06/05/2022, view: 600, Q&A: 0

This protocol describes a method to assess relative changes in the level of global protein synthesis in the preimplantation embryo using the Click-iT® Plus OPP Protein Synthesis Assays. In this assay, O-propargyl-puromycin (OPP), an analog of puromycin, is efficiently incorporated into the nascent polypeptide of newly translated proteins in

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Immunology

Protocol to Isolate Germinal Centers by Laser Microdissection Authors:  Farbod Bahreini, Markus Niebuhr, Julia Belde, Jürgen Westermann and Kathrin Kalies, date: 06/05/2022, view: 503, Q&A: 0

During adaptive immune responses, germinal centers (GC) appear as transient microstructures, in which antigen-specific B and T cells interact with each other. Because only the antigen-activated B and T cells, such as Plasmablasts or follicular T helper (Tfh) cells, are present in GC, the in depth-analysis of GC is of great interest. To identify

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Assessing the Presence of Hematopoietic Stem and Progenitor Cells in Mouse Spleen Authors:  Isabelle J. Marié, Lara Brambilla and David E. Levy, date: 06/05/2022, view: 644, Q&A: 0

Transplantation of hematopoietic material into recipient mice is an assay routinely used to determine the presence and function of hematopoietic stem and progenitor cells (HSPCs) in vivo. The principle of the method is to transplant donor cells being tested for HSPCs into a recipient mouse following bone marrow ablation and testing for

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Flow Cytometric Characterization of Macrophages Infected in vitro with Salmonella enterica Serovar Typhimurium Expressing Red Fluorescent Protein Authors:  Natascha Brigo, Christa Pfeifhofer-Obermair, Egon Demetz, Piotr Tymoszuk and Günter Weiss, date: 06/05/2022, view: 431, Q&A: 0

Macrophages are important for host defense against intracellular pathogens like Salmonella and can be differentiated into two major subtypes. M1 macrophages, which are pro-inflammatory and induce antimicrobial immune effector mechanisms, including the expression of inducible nitric oxide synthase (iNOS), and M2 macrophages, which exert

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Microbiology

A Modified Fluctuation Assay with a CAN1 Reporter in Yeast Authors:  Pengyao Jiang, Anja R. Ollodart and Maitreya J. Dunham, date: 06/05/2022, view: 518, Q&A: 0

Understanding the generation of mutations is fundamental to understanding evolution and genetic disease; however, the rarity of such events makes experimentally identifying them difficult. Mutation accumulation (MA) methods have been widely used. MA lines require serial bottlenecks to fix de novo mutations, followed by whole-genome sequencing.

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Plant Science

Investigation of Transposon DNA Methylation and Copy Number Variation in Plants Using Southern Hybridisation Authors:  Vivek Hari Sundar G. and P. V. Shivaprasad, date: 06/05/2022, view: 497, Q&A: 0

Plant genomes are pronouncedly enriched in repeat elements such as transposons. These repeats are epigenetically regulated by DNA methylation. Whole genome high-depth sequencing after bisulfite treatment remains an expensive and laborious method to reliably profile the DNA methylome, especially when considering large genomes such as in crops.

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