• 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.3930

Current Issue in 2021

Volume: 11 Issue: 4

Biochemistry

Expression and Purification of Yeast-derived GPCR, Gα and Gβγ Subunits for Structural and Dynamic Studies Authors:  Wenjie Zhao, Xudong Wang and Libin Ye, date: 02/20/2021, view: 953, Q&A: 0

In the last several years, as evidence of a surged number of GPCR-G complex structures, the expressions of GPCRs and G proteins for structural biology have achieved tremendous successes, mostly in insect and mammalian cell systems, resulting in more than 370 structures of over 70 GPCRs have been resolved. However, the challenge remains,

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Extraction and Quantification of Sphingolipids from Hemiptera Insects by Ultra-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry Authors:  Ni Wang, Xiaoxiao Shi, Chao Zhang, Wenwu Zhou and Zeng-Rong Zhu, date: 02/20/2021, view: 577, Q&A: 0

Sphingolipids are major structural components of endomembranes and have also been described as an intracellular second messenger involved in various biological functions in all eukaryotes and a few prokaryotes. Ceramides (Cer), the central molecules of sphingolipids, have been depicted in cell growth arrest, cell differentiation, and apoptosis.

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Tracing Nitrogen Metabolism in Mouse Tissues with Gas Chromatography-Mass Spectrometry Authors:  Rong Xu, Yekai Wang and Jianhai Du, date: 02/20/2021, view: 633, Q&A: 0

Nitrogen-containing metabolites including ammonia, amino acids, and nucleotides, are essential for cell metabolism, growth, and neural transmission. Nitrogen metabolism is tightly coordinated with carbon metabolism in the breakdown and biosynthesis of amino acids and nucleotides. Both nuclear magnetic resonance spectroscopy and mass spectrometry

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Biophysics

Live Cell FRET Analysis of the Conformational Changes of Human P-glycoprotein Authors:  Ryota Futamata, Noriyuki Kioka and Kazumitsu Ueda, date: 02/20/2021, view: 734, Q&A: 0

The molecular mechanisms of P-glycoprotein (P-gp; also known as MDR1 or ABCB1) have been mainly investigated using artificial membranes such as lipid-detergent mixed micelles, artificial lipid bilayers, and membrane vesicles derived from cultured cells. Although these in vitro experiments help illustrate details about the molecular mechanisms of

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Cell Biology

Bioorthogonal Labeling and Chemoselective Functionalization of Lung Extracellular Matrix Authors:  Zihan Ling, Yunhui Xing, Emily Reinoso Jacome, Shierly W. Fok and Xi Ren, date: 02/20/2021, view: 617, Q&A: 0

Decellularized extracellular matrix (ECM) biomaterials derived from native tissues and organs are widely used for tissue engineering and wound repair. To boost their regenerative potential, ECM biomaterials can be functionalized via the immobilization of bioactive molecules. To enable ECM functionalization in a chemoselective manner, we have

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Developmental Biology

Molecular and Phenotypic Characterization Following RNAi Mediated Knockdown in Drosophila Authors:  Saurabh Jayesh Kumar Mehta, Pradyumna A. Joshi and Ram Kumar Mishra, date: 02/20/2021, view: 646, Q&A: 0

Loss of function studies shed significant light on the involvement of a gene or gene product in different cellular processes. Short hairpin RNA (shRNA) mediated RNA interference (RNAi) is a classical yet straightforward technique frequently used to knock down a gene for assessing its function. Similar perturbations in gene expression can be

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Immunology

In vitro Measurement of Membrane Attack Complex in RPE Cells Authors:  Kelly Mulfaul and Sarah L. Doyle, date: 02/20/2021, view: 568, Q&A: 0

Initiation of the complement system results in the formation of a multiprotein pore termed the membrane attack complex (MAC, C5b-C9). MAC pores accumulate on a cell surface and can result in cell lysis. The retinal pigment epithelium (RPE) is a single monolayer of pigmented epithelial cells located at the posterior poll of the eye that forms the

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An Imaging Flow Cytometry Method to Measure Citrullination of H4 Histone as a Read-out for Neutrophil Extracellular Traps Formation Authors:  Emilia A. Barbu, Venina M. Dominical, Laurel Mendelsohn and Swee Lay Thein, date: 02/20/2021, view: 577, Q&A: 0

The formation of neutrophil extracellular traps (NETs) is thought to play a critical role in infections and propagating sterile inflammation. Histone citrullination is an essential and early step in NETs formation, detectable prior to the formation of the hallmark extracellular DNA-scaffolded strands. In addition to the classical microscopy

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Microbiology

Rapid Genome Engineering of Pseudomonas Assisted by Fluorescent Markers and Tractable Curing of Plasmids Authors:  Daniel C. Volke, Nicolas T. Wirth and Pablo I. Nikel, date: 02/20/2021, view: 1221, Q&A: 0

Precise genome engineering has become a commonplace technique for metabolic engineering. Also, insertion, deletion and alteration of genes and other functional DNA sequences are essential for understanding and engineering cells. Several techniques have been developed to this end (e.g., CRISPR/Cas-assisted methods, homologous recombination, or λ

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Molecular Biology

RI-SEC-seq: Comprehensive Profiling of Nonvesicular Extracellular RNAs with Different Stabilities Authors:  Juan Pablo Tosar, Fabiana Gámbaro, Mauricio Castellano and Alfonso Cayota, date: 02/20/2021, view: 775, Q&A: 0

Exosomes and other extracellular vesicles (EVs) are considered the main vehicles transporting RNAs in extracellular samples, including human bodily fluids. However, a major proportion of extracellular RNAs (exRNAs) do not copurify with EVs and remain in ultracentrifugation supernatants of cell-conditioned medium or blood serum. We have observed

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Plant Science

Phytophthora infestans (Late blight) Infection Assay in a Detached Leaf of Potato Authors:  Hari S. Karki and Dennis A. Halterman, date: 02/20/2021, view: 674, Q&A: 0

Phytophthora infestans is a hemibiotroph oomycete that primarily infects potato and tomato. It infects stems, leaves, and tubers and fruits of potato and tomato. High throughput and reproducible infection assays are prerequisites to find sources of resistance in any crop. In this protocol, we describe a detached leaf assay (DLA) for conducting the

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Stem Cell

Generation of the Compression-induced Dedifferentiated Adipocytes (CiDAs) Using Hypertonic Medium Authors:  Yiwei Li, Angelo S. Mao, Bo Ri Seo, Xing Zhao, Satish Kumar Gupta, Maorong Chen, Yu Long Han, Ting-Yu Shih, David J. Mooney and Ming Guo, date: 02/20/2021, view: 476, Q&A: 0

Current methods to obtain mesenchymal stem cells (MSCs) involve sampling, culturing, and expanding of primary MSCs from adipose, bone marrow, and umbilical cord tissues. However, the drawbacks are the limited numbers of total cells in MSC pools, and their decaying stemness during in vitro expansion. As an alternative resource, recent ceiling

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Systems Biology

EmPC-seq: Accurate RNA-sequencing and Bioinformatics Platform to Map RNA Polymerases and Remove Background Error Authors:  Yuqing Wang, Tin Hang Chong, Ilona Christy Unarta, Xinzhou Xu, Gianmarco D. Suarez, Jiguang Wang, John T. Lis, Xuhui Huang and Peter Pak-Hang Cheung, date: 02/20/2021, view: 438, Q&A: 0

Transcription errors can substantially affect metabolic processes in organisms by altering the epigenome and causing misincorporations in mRNA, which is translated into aberrant mutant proteins. Moreover, within eukaryotic genomes there are specific Transcription Error-Enriched genomic Loci (TEELs) which are transcribed by RNA polymerases with

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