• 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.4367

Past Issues in 2022

Volume: 12 Issue: 7

Biochemistry

A SYBR Gold-based Label-free in vitro Dicing Assay Authors:  Qian Wang, Yan Xue, Laixing Zhang, Zhenhui Zhong, Suhua Feng, Changshi Wang, Lifan Xiao, Zhenlin Yang, C. Jake Harris, Zhe Wu, Jixian Zhai, Maojun Yang, Sisi Li, Steven E. Jacobsen and Jiamu Du, date: 04/05/2022, view: 577, Q&A: 0

In Arabidopsis, DICER-LIKE PROTEIN 3 (DCL3) cuts the substrate pre-siRNA into a product siRNA duplex, encompassing one 23-nt strand and one 24-nt strand. To monitor the separation of the siRNA duplex with only 1-nt difference, we developed this protocol to evaluate the in vitro dicing activity of DCL3. The method can be applied for measuring

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Biological Engineering

Activation of Mitochondrial Ca2+ Oscillation and Mitophagy Induction by Femtosecond Laser Photostimulation Authors:  Xiaoying Tian and Hao He, date: 04/05/2022, view: 319, Q&A: 0

Ultra-precise stimulation solely to individual mitochondria, without any influence to the whole cell, is quite difficult by traditional biochemical reagents. In mitophagy research, the mitochondria and even the whole cell usually suffer irreversible and great damage caused by treatment with potent chemicals. In this protocol, we present the

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Quantitative and Anatomical Imaging of Human Skin by Noninvasive Photoacoustic Dermoscopy Authors:  Zhiyang Wang, Fei Yang, Wuyu Zhang and Sihua Yang, date: 04/05/2022, view: 370, Q&A: 0

Imaging plays a vital role in the diagnosis and treatment of skin diseases. However, pure optical imaging technique is limited to the visualization of superficial skin tissues. Ultrasonic imaging technique can detect deep tissues, but it lacks detailed information on microscopic pathological structures. Photoacoustic imaging is an advanced

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Ex-vivo Skin Permeability Tests of Nanoparticles for Microscopy Imaging Authors:  Nethanel Friedman, Sharon Merims, Jhonatan Elia and Ofra Benny, date: 04/05/2022, view: 372, Q&A: 0

Delivery of drugs through the skin is a major challenge in the field of drug delivery systems. Quantification of materials, and specifically nanoparticles, within the skin layers is essential for the development of advanced topical and transdermal delivery systems. We have developed a technique for ex-vivo segmentation and evaluation of human

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Cancer Biology

Image-based Quantification of Macropinocytosis Using Dextran Uptake into Cultured Cells Authors:  Anh H. Le and Laura M. Machesky, date: 04/05/2022, view: 519, Q&A: 0

Macropinocytosis is an evolutionarily conserved process, which is characterized by the formation of membrane ruffles and the uptake of extracellular fluid. We recently demonstrated a role for CYFIP-related Rac1 Interactor (CYRI) proteins in macropinocytosis. High-molecular weight dextran (70kDa or higher) has generally been used as a marker for

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Conditional Human BRD4 Knock-In Transgenic Mouse Genotyping and Protein Isoform Detection Authors:  Michael Paul Lewis, Shwu-Yuan Wu and Cheng-Ming Chiang, date: 04/05/2022, view: 425, Q&A: 0

Bromodomain-containing protein 4 (BRD4) is an acetyl-lysine reader protein and transcriptional regulator implicated in chromatin dynamics and cancer development. Several BRD4 isoforms have been detected in humans with the long isoform (BRD4-L, aa 1-1,362) playing a tumor-suppressive role and a major short isoform (BRD4-S, aa 1-722) having

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Cell Biology

In vivo Ca2+ Imaging in Mouse Salivary Glands Authors:  Takahiro Takano and David I. Yule, date: 04/05/2022, view: 668, Q&A: 0

Changes in intracellular calcium drive exocrine cell activity. In the salivary gland, acetylcholine released from parasympathetic neurons mobilizes endoplasmic reticulum calcium stores in acinar cells, which consequently initiates saliva secretion. However, our understanding of the signaling cascade is mainly based on ex vivo studies

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Developmental Biology

Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice Authors:  Hiroki Ueharu, Jingwen Yang, Yoshihiro Komatsu and Yuji Mishina, date: 04/05/2022, view: 456, Q&A: 0

Craniofacial anomalies (CFA) are a diverse group of deformities, which affect the growth of the head and face. Dysregulation of cranial neural crest cell (NCC) migration, proliferation, differentiation, and/or cell fate specification have been reported to contribute to CFA. Understanding of the mechanisms through which cranial NCCs contribute

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Immunology

Identification of SARS-CoV-2 Neutralizing Antibody with Pseudotyped Virus-based Test on HEK-293T hACE2 Cells Authors:  Constant Gillot, Julien Favresse, Vincent Maloteau, Jean-Michel Dogné and Jonathan Douxfils, date: 04/05/2022, view: 393, Q&A: 0

Neutralizing antibodies (NAbs) are of particular importance because they can prevent binding of the receptor binding domain (RBD) of the spike protein (S protein) to the angiotensin-converting enzyme 2 (ACE2) receptor present at the surface of human cells, preventing virus entry into the host cells. The gold standard method for detection of

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A Mouse Infection Model with a Wildtype Salmonella enterica Serovar Typhimurium Strain for the Analysis of Inflammatory Innate Immune Cells Authors:  Christa Pfeifhofer-Obermair, Natascha Brigo, Piotr Tymoszuk and Günter Weiss, date: 04/05/2022, view: 339, Q&A: 0

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a Gram-negative, facultative intracellular bacterium, which causes gastrointestinal disorders in humans, and systemic, typhoid fever-like infections in mice. Our current knowledge regarding the involvement of cellular and humoral immunity in the defense from S. Typhimurium infections is

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Medicine

Preparation, Characterization, and Cell Uptake of PLGA/PLA-PEG-FA Nanoparticles Authors:  Heng Yang, Junsik Sung, Zahra Alghoul and Didier Merlin, date: 04/05/2022, view: 349, Q&A: 0

Oral administration of colon-targeting nanoformulations holds many advantages over the systemic delivery of free drugs, or traditional nontargeting formulations in the treatment of ulcerative colitis (UC). Currently, the most conventional method for constructing colon-targeting drug delivery systems (DDS) is by integrating the biocompatible

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Molecular Biology

Plasmid and Sequencing Library Preparation for CRISPRi Barcoded Expression Reporter Sequencing (CiBER-seq) in Saccharomyces cerevisiae Authors:  Ryan Y Muller, Zuriah A Meacham and Nicholas T Ingolia, date: 04/05/2022, view: 679, Q&A: 0

Genetic networks regulate nearly all biological processes, including cellular differentiation, homeostasis, and immune responses. Determining the precise role of each gene within a regulatory network can explain its overall, integrated function, and pinpoint mechanisms underlying misregulation in disease states. Transcriptional reporter assays are

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A Rapid FRET Real-Time PCR Protocol for Simultaneous Quantitative Detection and Discrimination of Human Plasmodium Parasites Authors:  Renate Schneider, Aline Lamien-Meda, Herbert Auer, Ursula Wiedermann-Schmidt, Peter L. Chiodini and Julia Walochnik, date: 04/05/2022, view: 316, Q&A: 0

Malaria is the most important parasitic disease worldwide, and accurate diagnosis and treatment without delay are essential for reducing morbidity and mortality, especially in P. falciparum malaria. Real-time PCR is highly sensitive and highly specific, therefore an excellent diagnostic tool for laboratory detection and species-specific

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Neuroscience

Multiplexing Thermotaxis Behavior Measurement in Caenorhabditis elegans Authors:  Stephan Raiders, Mason Klein and Aakanksha Singhvi, date: 04/05/2022, view: 686, Q&A: 0

Thermotaxis behaviors in C. elegans exhibit experience-dependent plasticity of thermal preference memory. This behavior can be assayed either at population level, on linear temperature gradients, or at the individual animal level, by radial isothermal or microfluidic tracking of orientation. These behaviors are low-throughput as well as

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Stimulus-induced Robust Narrow-band Gamma Oscillations in Human EEG Using Cartesian Gratings Authors:  Dinavahi V. P. S. Murty and Supratim Ray, date: 04/05/2022, view: 316, Q&A: 0

Stimulus-induced narrow-band gamma oscillations (20–70 Hz) are induced in the visual areas of the brain when particular visual stimuli, such as bars, gratings, or full-screen hue, are shown to the subject. Such oscillations are modulated by higher cognitive functions, like attention, and working memory, and have been shown to be abnormal in

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Plant Science

In vitro Auto- and Substrate-Ubiquitination Assays Authors:  Hye Lin Park, Han Yong Lee and Gyeong Mee Yoon, date: 04/05/2022, view: 443, Q&A: 0

The precise regulation of the homeostasis of the cellular proteome is critical for the appropriate growth and development of plants. It also allows the plants to respond to various environmental stresses, by modulating their biochemical and physiological aspects in a timely manner. Ubiquitination of cellular proteins is one of the major

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