Volume: 9 Issue: 24 —BIO-PROTOCOL

Biochemistry

Semi-quantitative Determination of Protein Expression Using Immunohistochemistry Staining and Analysis: An Integrated Protocol Authors: Alexandra R Crowe and Wei Yue, date: 12/20/2019, view: 1984, Q&A: 0

Semi-quantitative IHC is a powerful method for investigating protein expression and localization within tissues. The semi-quantitative immunohistochemistry (IHC) involves using software such as free software ImageJ Fiji to conduct deconvolution and downstream analysis. Currently, there is lack of an integrated protocol that includes a detailed ...

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Cell Biology

Analysis of EGF Receptor Endocytosis Using Fluorogen Activating Protein Tagged Receptor Authors: Mireia Perez Verdaguer, Mads B. Larsen, Marcel P. Bruchez, Simon C. Watkins and Alexander Sorkin, date: 12/20/2019, view: 1352, Q&A: 0

Functional activities of many transmembrane proteins are controlled by their endocytosis. One of the most studied experimental models is the epidermal growth factor (EGF) receptor (EGFR). However, endocytic trafficking of EGFR has been predominantly analyzed using labeled EGF, whereas quantitative analyses of the endocytosis of the receptor itself ...

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Immunology

Lysosome Targeting RedGreen-assay: Selective Autophagy Sensing Assay for Mammalian Cells Authors: Victoria Riccio, Miluska Vissa, Angus McQuibban and Peter Kijun Kim, date: 12/20/2019, view: 1540, Q&A: 0

The process of autophagy is an essential cellular mechanism, required to maintain general cell health through the removal of dysfunctional organelles, such as the ER, peroxisomes and mitochondria, as well as protein aggregates, and bacteria. Autophagy is an extremely dynamic process, and tools are constantly being developed to study the various ...

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cDNA Display Mediated Immuno-PCR (cD-IPCR): A Novel PCR-based Antigen Detection Method Authors: Chathuni Jayathilake, Takuya Terai and Naoto Nemoto, date: 12/20/2019, view: 1645, Q&A: 0

Immuno-PCR (IPCR) is a powerful method in antigen detection where a PCR-amplifiable DNA reporter is conjugated to a specific antibody or an aptamer for the target molecule. In the development and application of IPCR, successful conjugation of a protein (an antibody) with a reporter DNA becomes challenging. To address this issue, we recently ...

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Propagation and Purification of Chlamydia trachomatis Serovar L2 Transformants and Mutants Authors: Robert Faris and Mary M. Weber, date: 12/20/2019, view: 1041, Q&A: 0

Chlamydia trachomatis (C.t.) is an obligate intracellular pathogen that cannot be cultured axenically and must be propagated within eukaryotic host cells. There are at least 15 distinct chlamydial serovariants that belong to 2 major biovars commonly referred to as trachoma and lymphogranuloma venereum (LGV). The invasive ...

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Isolation and Stimulation of Peritoneal Macrophages with Apoptotic Jurkat Cells to Produce IL-10 Authors: Mei Song and Xiaojing Ma, date: 12/20/2019, view: 1166, Q&A: 0

Clearance of apoptotic cells by macrophages is critical to ensuring cellular homeostasis and suppression of autoimmunity. Macrophage recognition of apoptotic cells triggers an anti-inflammatory response, which is mediated by the release of IL-10, TGF-β etc. with concurrent inhibition of pro-inflammatory cytokines (such as TNFα, IL-12, ...

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Microbiology

Preparation of Red Palm Weevil Rhynchophorus Ferrugineus (Olivier) (Coleoptera: Dryophthoridae) Germ-free Larvae for Host-gut Microbes Interaction Studies Authors: Abrar Muhammad, Prosper Habineza, Youming Hou and Zhanghong Shi, date: 12/20/2019, view: 1249, Q&A: 0

Red palm weevil (RPW), Rhynchophorus ferrugineus Olivier, is a devastating pest of palm trees worldwide. RPW gut is colonized by diverse bacterial species which profoundly influence host development and nutritional metabolism. However, the molecular mechanisms behind the interactions between RPW and its gut microbiota remain mostly ...

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Preparation of Actinoplanes missouriensis Zoospores and Assay for Their Adherence to Solid Surfaces Authors: Takeaki Tezuka, Daisuke Nakane, Tomohiro Kimura and Yasuo Ohnishi, date: 12/20/2019, view: 760, Q&A: 0

Spherical zoospores of a rare actinomycete, Actinoplanes missouriensis, adhere to various hydrophobic solid surfaces by means of type IV pili. The purpose of this protocol is to provide detailed descriptions of the preparation of A. missouriensis zoospores and an assay for the adhesion of the zoospores to solid surfaces. This ...

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Protocol for Ribosome Profiling in Bacteria Authors: Fuad Mohammad and Allen R. Buskirk, date: 12/20/2019, view: 2109, Q&A: 0

Ribosome profiling provides information on the position of ribosomes on mRNA on a genomic scale. Although this information is often used to detect changes in gene expression under different conditions, it also has great potential for yielding insight into the mechanism and regulation of protein synthesis itself. First developed in yeast, ribosome ...

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Neuroscience

Myelin Oligodendrocyte Glycoprotein 35-55 (MOG 35-55)-induced Experimental Autoimmune Encephalomyelitis: A Model of Chronic Multiple Sclerosis Authors: Sakie Miyamura, Nagisa Matsuo, Kazuki Nagayasu, Hisashi Shirakawa and Shuji Kaneko, date: 12/20/2019, view: 1492, Q&A: 0

Multiple sclerosis (MS) is the common demyelinating disease of human central nervous system. Among mouse models available to study MS, including the cuprizone application and lysolecithin-injection models, experimental autoimmune encephalomyelitis (EAE) model is widely used so that chronic EAE model of C57BL/6J can reflect the autoimmune ...

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Semi-automated Model to Accurately Counting Sympathetic Nervous Fibers Authors: Dennis Bleck, Lkham Erdene-Byambadoo, Ralph Brinks, Matthias Schneider and Georg Pongratz, date: 12/20/2019, view: 947, Q&A: 0

In recent years, the role of sympathetic nervous fibers in chronic inflammation has become increasingly evident. At the onset of inflammation, sympathetic activity is increased in the affected tissue. However, sympathetic fibers are largely absent from chronically inflamed tissues. Apparently, there is a very dynamic relationship between ...

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Plant Science

Analysis of Monosaccharides from Arabidopsis Seed Mucilage and Whole Seeds Using HPAEC-PAD Authors: Gillian H. Dean, Kresimir Sola, Faride Unda, Shawn D. Mansfield and George W. Haughn, date: 12/20/2019, view: 1347, Q&A: 0

Arabidopsis seed coat epidermal cells deposit a significant quantity of mucilage, composed of the cell wall components pectin, hemicellulose, and cellulose, into the apoplast during development. When mature seeds are hydrated, mucilage extrudes to form a gelatinous capsule around the seed. Determining the monosaccharide composition of ...

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Stem Cell

Retina Injury and Retina Tissue Preparation to Study Regeneration in Zebrafish Authors: Poonam Sharma and Rajesh Ramachandran, date: 12/20/2019, view: 1268, Q&A: 0

Unlike mammals, primitive vertebrates have immense capability to regenerate almost all of their organs including the central nervous system. Among primitive organisms, zebrafish have been extensively used as a model system for regeneration studies. The retina is a part of the central nervous system and mammals lack the potential to repair any ...

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In vitro Differentiation of Human iPSC-derived Retinal Pigment Epithelium Cells (iPSC-RPE) Authors: Agnieszka D’Antonio-Chronowska, Matteo D’Antonio and Kelly A. Frazer, date: 12/20/2019, view: 1863, Q&A: 0

Induced Pluripotent Stem Cells (iPSCs) serve as an excellent model system for studying the molecular underpinnings of tissue development. Human iPSC-derived retinal pigment epithelium (iPSC-RPE) cells have fetal-like molecular profiles. Hence, biobanks like iPSCORE, which contain iPSCs generated from hundreds of individuals, are an invaluable ...

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Past Issues in 2019

Volume: 9 Issue: 24

Biochemistry

Cell Biology

Immunology

Microbiology

Neuroscience

Plant Science

Stem Cell