• 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.4422

Past Issues in 2022

Volume: 12 Issue: 10

Biological Engineering

Preparation of Cas9 Ribonucleoproteins for Genome Editing Authors:  Sheng-Wei Lin, Viet Quoc Nguyen and Steven Lin, date: 05/20/2022, view: 1333, Q&A: 0

Genome editing by the delivery of pre-assembled Cas9 ribonucleoproteins (Cas9 RNP) is an increasingly popular approach for cell types that are difficult to manipulate genetically by the conventional plasmid and viral methods. Cas9 RNP editing is robust, precise, capable of multiplexing, and free of genetic materials. Its transient presence in

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Immunomagnetic Isolation and Enrichment of Microvascular Endothelial Cells from Human Adipose Tissue Authors:  Jeremy A. Antonyshyn, Vienna Mazzoli, Meghan J. McFadden, Anthony O. Gramolini, Stefan O. P. Hofer, Craig A. Simmons and J. Paul Santerre, date: 05/20/2022, view: 866, Q&A: 0

Human adipose tissue-resident microvascular endothelial cells are not only garnering attention for their emergent role in the pathogenesis of obesity-related metabolic disorders, but are also of considerable interest for vascular tissue engineering due, in part, to the abundant, accessible, and uniquely dispensable nature of the tissue. Here, we

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Biophysics

Purification of Mitochondrial Ribosomal Complexes from Trypanosoma cruzi and Leishmania tarentolae for Cryo-EM Analysis Authors:  Stéphanie Durrieu-Gaillard, Marie Sissler and Yaser Hashem, date: 05/20/2022, view: 917, Q&A: 1

Kinetoplastids are unicellular eukaryotic parasites responsible for human pathologies such as Chagas disease, sleeping sickness or Leishmaniasis, caused by Trypanosoma cruzi, Trypanosoma brucei, and various Leishmania spp., respectively. They harbor a single large mitochondrion that is essential for the survival of the parasite. Interestingly,

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Cancer Biology

A Flow Cytometry-Based Method for Analyzing DNA End Resection in G0- and G1-Phase Mammalian Cells Authors:  Bo-Ruei Chen, Jessica K. Tyler and Barry P. Sleckman, date: 05/20/2022, view: 804, Q&A: 0

DNA double strand breaks (DSBs) constantly arise in cells during normal cellular processes or upon exposure to genotoxic agents, and are repaired mostly by homologous recombination (HR) and non-homologous end joining (NHEJ). One key determinant of DNA DSB repair pathway choice is the processing of broken DNA ends to generate single strand DNA

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A Highly Sensitive Method to Efficiently Profile the Histone Modifications of FFPE Samples Authors:  Linxuan Zhao, Vamsi Krishna Polavarapu, Ram Prakash Yadav, Pengwei Xing and Xingqi Chen, date: 05/20/2022, view: 758, Q&A: 0

The majority of biopsies in both basic research and translational cancer studies are preserved in the format of archived formalin-fixed paraffin-embedded (FFPE) samples. Profiling histone modifications in archived FFPE tissues is critically important to understand gene regulation in human disease. The required input for current genome-wide

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Cell Biology

Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat Authors:  Huiliang Zhang and Peter S. Rabinovitch, date: 05/20/2022, view: 895, Q&A: 0

The isolation of intact single adult cardiomyocytes from model animals, mouse and rat, is an essential tool for cardiac molecular and cellular research. While several methods are reported for adult mouse cardiomyocyte isolation, the viability and yield of the isolated cells have been variable. Here, we describe step-by-step protocols for high

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CRISPR/Cas9 Gene Editing of HeLa Cells to Tag Proteins with mNeonGreen Authors:  Sachin Surve and Alexander Sorkin, date: 05/20/2022, view: 1031, Q&A: 0

Subcellular localization dynamics of proteins involved in signal transduction processes is crucial in determining the signaling outcome. However, there is very limited information about the localization of endogenous signaling proteins in living cells. For example, biochemical mechanisms underlying the signaling pathway from epidermal growth

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Developmental Biology

Preparation of a Single Cell Suspension from the Murine Iridocorneal Angle Authors:  Benjamin R. Thomson and Susan E. Quaggin, date: 05/20/2022, view: 591, Q&A: 0

Single cell RNA sequencing is a powerful tool that can be used to identify distinct cell types and transcriptomic differences within complex tissues. It has proven to be especially useful in tissues of the eye, where investigators have identified novel cell types within the retina, anterior chamber, and iridocorneal angle and explored

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Immunology

Protocol to Induce Follicular T Helper Cells, Germinal Centers, and Skin Lesions in Mouse Models for Skin Blistering Diseases Authors:  Farbod Bahreini, Markus Niebuhr, Julia Belde, Katja Bieber, Juergen Westermann and Kathrin Kalies, date: 05/20/2022, view: 856, Q&A: 0

Autoreactive T cells in autoantibody-mediated autoimmune diseases can be divided into two major subsets: (i) follicular T helper cells (Tfh) that provide T cell help in germinal centers (GC) and (ii) effector T (Teff) cells that immigrate into peripheral tissue sites such as the skin and mediate local inflammation. To study the sequence of events

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Microbiology

Flow Cytometry-based Measurement of Reactive Oxygen Species in Cyanobacteria Authors:  Soumila Mondal and Shailendra P. Singh, date: 05/20/2022, view: 638, Q&A: 0

Cyanobacteria are Gram-negative oxygen-producing photosynthetic bacteria that are useful in the pharmaceutical and biofuel industries. Monitoring of oxidative stress under fluctuating environmental conditions is important for determining the fitness, survival, and growth of cyanobacteria in the laboratory as well as in large scale cultivation

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Purification of the Bacterial Amyloid “Curli” from Salmonella enterica Serovar Typhimurium and Detection of Curli from Infected Host Tissues Authors:  Murugesan Sivaranjani, Elizabeth G Hansen, Sumudu R. Perera, Pamela A. Flores, Çagla Tükel and Aaron P. White, date: 05/20/2022, view: 465, Q&A: 0

Microbiologists are learning to appreciate the importance of “functional amyloids” that are produced by numerous bacterial species and have impacts beyond the microbial world. These structures are used by bacteria to link together, presumably to increase survival, protect against harsh conditions, and perhaps to influence cell-cell

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Neuroscience

Vestibular Organ Dissection and Whole-Mount Immunolabeling in Mouse Authors:  Timothy S. Balmer and Laurence O. Trussell, date: 05/20/2022, view: 424, Q&A: 0

The vestibular sensory apparatus contained in the inner ear is a marvelous evolutionary adaptation for sensing movement in 3 dimensions and is essential for an animal’s sense of orientation in space, head movement, and balance. Damage to these systems through injury or disease can lead to vertigo, Meniere’s disease, and other disorders

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Identification and Monitoring of Nucleotide Repeat Expansions Using Southern Blotting in Drosophila Models of C9orf72 Motor Neuron Disease and Frontotemporal Dementia Authors:  Joanne L. Sharpe, Nikki S. Harper and Ryan J. H. West, date: 05/20/2022, view: 416, Q&A: 0

Repeat expansion diseases, including fragile X syndrome, Huntington’s disease, and C9orf72-related motor neuron disease and frontotemporal dementia, are a group of disorders associated with polymorphic expansions of tandem repeat nucleotide sequences. These expansions are highly repetitive and often hundreds to thousands of repeats in

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Plant Science

In vitro Assays to Evaluate Specificity and Affinity in Protein-phospholipid Interactions Authors:  Hendry Susila, Snježana Jurić, Katarzyna Gawarecka, Kyung Sook Chung, Suhyun Jin, Soo-Jin Kim, Zeeshan Nasim, Geummin Youn and Ji Hoon Ahn, date: 05/20/2022, view: 655, Q&A: 0

Protein–lipid interactions play important roles in many biological processes, including metabolism, signaling, and transport; however, computational and structural analyses often fail to predict such interactions, and determining which lipids participate in these interactions remains challenging. In vitro assays to assess the physical

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Stem Cell

Optimized CRISPR-Cas9-based Strategy for Complex Gene Targeting in Murine Embryonic Stem Cells for Germline Transmission Authors:  Thomas J. O'Neill, Daniel Krappmann and Andreas Gewies, date: 05/20/2022, view: 815, Q&A: 0

Although CRISPR-Cas9 genome editing can be performed directly in single-cell mouse zygotes, the targeting efficiency for more complex modifications such as the insertion of two loxP sites, multiple mutations in cis, or the precise insertion or deletion of longer DNA sequences often remains low (Cohen, 2016). Thus, targeting and validation of

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