• 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.4465

Current Issue in 2022

Volume: 12 Issue: 15

Biochemistry

Evaluation of Urine Proteins by Capillary Electrophoresis Authors:  Paula F. Navarro, Laura Gil and Salceda Fernández-Barredo, date: 08/05/2022, view: 170, Q&A: 0

Capillary electrophoresis (CE) is a laboratory method usually used to separate proteins in body fluids such as serum, cerebrospinal fluid, or urine. Separation of proteins in urine can have clinical applications for evaluating samples from healthy dogs and dogs with proteinuria in a qualitative way, which would not be possible with gel

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Purification of Crimean Congo Hemorrhagic Fever Virus (CCHFV) Nucleocapsid Protein Using Detergent Gradient and Free Thawing Authors:  Austin Royster and Sheema Mir, date: 08/05/2022, view: 122, Q&A: 0

Protein aggregation remains a major challenge in the purification of recombinant proteins in both eukaryotic and prokaryotic expression systems. One such protein is the nucleocapsid protein of Crimean Congo Hemorrhagic fever virus (CCHFV), which has high aggregation tendency and rapidly precipitates upon purification by NiNTA chromatography. Using

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Biophysics

Visualization, Quantification, and Modeling of Endogenous RNA Polymerase II Phosphorylation at a Single-copy Gene in Living Cells Authors:  Linda S. Forero-Quintero, William Raymond, Brian Munsky and Timothy J. Stasevich, date: 08/05/2022, view: 367, Q&A: 0

In eukaryotic cells, RNA Polymerase II (RNAP2) is the enzyme in charge of transcribing mRNA from DNA. RNAP2 possesses an extended carboxy-terminal domain (CTD) that gets dynamically phosphorylated as RNAP2 progresses through the transcription cycle, therefore regulating each step of transcription from recruitment to termination. Although RNAP2

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Cancer Biology

CRISPR/Cas9-mediated Gene Knockout Followed by Negative Selection Leads to a Complete TCR Depletion in orthoCAR19 T Cells Authors:  Qian Zhang, Jingyi Yang, Eric Nigel Ebenezer Anand Manoharan, Alvin B. Yu and Michael C. Milone, date: 08/05/2022, view: 230, Q&A: 0

Genome-editing technologies, especially CRISPR (clustered regularly interspaced short palindrome repeats)/Cas9 (CRISPR-associated protein 9), endows researchers the ability to make efficient, simple, and precise genomic DNA changes in many eukaryotic cell types. CRISPR/Cas9-mediated efficient gene knockout holds huge potential to improve the

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Computational Biology and Bioinformatics

TRACES: a Freely Accessible, Semi-automated Pipeline for Detection, Tracking, and Quantification of Fluorescently Labeled Cellular Structures Authors:  Xueer Jiang, Linhao Jiang and Li-En Jao, date: 08/05/2022, view: 118, Q&A: 0

Subcellular structures exhibit diverse behaviors in different cellular processes, including changes in morphology, abundance, and relative spatial distribution. Faithfully tracking and quantifying these changes are essential to understand their functions. However, most freely accessible methods lack integrated features for tracking multiple

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Drug Discovery

Generation of a Human Conditionally Immortalized Cell-based Multicellular Spheroidal Blood-Brain Barrier Model for Permeability Evaluation of Macromolecules Authors:  Ryuto Isogai, Hanae Morio, Ayaka Okamoto, Keita Kitamura and Tomomi Furihata, date: 08/05/2022, view: 173, Q&A: 0

There is an urgent need for the development of brain drug delivery carriers based on middle-sized or macromolecules, to which in vitro blood-brain barrier (BBB) models are expected to contribute significantly through evaluation of BBB permeability. As part of efforts to develop such models, we have been working on human conditionally

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Immunology

Cost Effective Method for gDNA Isolation from the Cecal Content and High Yield Procedure for RNA Isolation from the Colonic Tissue of Mice Authors:  Sohini Mukhopadhyay and Palok Aich, date: 08/05/2022, view: 185, Q&A: 0

Microbiome studies are quickly gaining momentum. Since most of the resident microbes (consisting of bacteria, fungi, and viruses) are difficult to culture, sequencing the microbial genome is the method of choice to characterize them. It is therefore important to have efficient methodology for gDNA isolation of gut microbes. Mouse models are widely

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Microbiology

Cryptococcus neoformans Virulence Assay Using a Galleria mellonella Larvae Model System Authors:  Piotr R. Stempinski, Daniel F. Q. Smith and Arturo Casadevall, date: 08/05/2022, view: 283, Q&A: 0

Cryptococcus neoformans is a human pathogenic fungus that can cause pulmonary infections and meningitis in both immunocompromised and otherwise healthy individuals. Limited treatment options and a high mortality rate underlie the necessity for extensive research of the virulence of C. neoformans. Here we describe a detailed protocol for using the

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Molecular Biology

DSP-crosslinking and Immunoprecipitation to Isolate Weak Protein Complex Authors:  Kotaro Akaki, Takashi Mino and Osamu Takeuchi, date: 08/05/2022, view: 145, Q&A: 0

Detecting protein-protein interactions (PPIs) is one of the most used approaches to reveal the molecular regulation of protein of interests (POIs). Immunoprecipitation of POIs followed by mass spectrometry or western blot analysis enables us to detect co-precipitated POI-binding proteins. However, some binding proteins are lost during cell lysis

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Neuroscience

A Step-by-step Protocol for Obtaining Mature Microglia from Mice Authors:  Min-Jung Yoo and Min-Soo Kwon, date: 08/05/2022, view: 321, Q&A: 0

In mice, microglial precursors in the yolk sac migrate to the brain parenchyma through the head neuroepithelial layer between embryonic days 8.5 (E8.5)–E16.5 and acquire their unique identity with a ramified form. Based on the microglial developmental process, we dissected the neuroepithelial layer (NEL) of E13.5 mice, which is composed

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