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324503
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关键词:'324503'
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Christos D Georgiou et al.
Redox biology, 17, 236-245 (2018-05-05)
A new fluorometric assay is presented for the ultrasensitive quantification of total protein carbonyls, and is based on their specific reaction with rhodamine B hydrazide (RBH), and the production of a protein carbonyl-RBH hydrazone the fluorescence of which (at ex/em
Gabriel Mendoza-Rojas et al.
STAR protocols, 2(4), 100899-100899 (2021-11-13)
Here, we describe a detailed step-by-step protocol for the expression, purification, quantification, and activity determination of key enzymes for molecular detection of pathogens. Based on previous reports, we optimized the protocol for LbCas12a, Taq DNA polymerase, M-MLV reverse transcriptase, and
Ana Paula Zen Petisco Fiore et al.
Bio-protocol, 13(18), e4822-e4822 (2023-09-27)
Eukaryotic cells have different types of proteasomes that differ in size. The smallest proteolytically active particle is the 20S proteasome, which degrades damaged and oxidized proteins; the most common larger particle is the 26S proteasome, which degrades ubiquitylated proteins. The
Qasim Ali et al.
MethodsX, 6, 259-264 (2019-02-23)
Extraction of high-quality DNA from Gossypium (Cotton) species is notoriously difficult due to high contents of polysaccharides, quinones and polyphenols other secondary metabolites. Here, we describe a simple, rapid and modified procedure for high-quality DNA extraction from cotton, which is
Anne A M J Becker et al.
PloS one, 10(4), e0123933-e0123933 (2015-04-24)
Understanding the symbiotic relationship between gut microbes and their animal host requires characterization of the core microbiota across populations and in time. Especially in captive populations of endangered wildlife species such as the cheetah (Acinonyx jubatus), this knowledge is a
Camilla Molinari et al.
Current protocols, 2(8), e523-e523 (2022-08-24)
The blast fungus, Magnaporthe oryzae, is a devastating plant pathogen that threatens global food security. The social and economic importance of blast disease has contributed to this filamentous fungus becoming a model organism for the study of host-pathogen interactions. Availability
Nike Walther et al.
The Journal of cell biology, 217(7), 2309-2328 (2018-04-11)
The two Condensin complexes in human cells are essential for mitotic chromosome structure. We used homozygous genome editing to fluorescently tag Condensin I and II subunits and mapped their absolute abundance, spacing, and dynamic localization during mitosis by fluorescence correlation
Roberto Alcántara et al.
Cell reports methods, 1(7), 100093-100093 (2021-10-27)
Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced
Patryk Ludzia et al.
Structure (London, England : 1993), 29(9), 1014-1028 (2021-04-30)
The kinetochore is the macromolecular machinery that drives chromosome segregation by interacting with spindle microtubules. Kinetoplastids (such as Trypanosoma brucei), a group of evolutionarily divergent eukaryotes, have a unique set of kinetochore proteins that lack any significant homology to canonical
Alex Martínez-Sabadell et al.
STAR protocols, 3(4), 101712-101712 (2022-11-02)
Immunotherapy has revolutionized cancer treatment, but preclinical models are required to understand immunotherapy resistance mechanisms underlying patient relapse. This protocol describes how to generate an acquired resistance humanized in vivo model to immunotherapies in patient-derived xenografts (PDX). We detail steps to
Yuzuru Taguchi et al.
Bio-protocol, 4(11) (2014-06-05)
Here, we introduce the protocol for small-scale and simple subcellular fractionation used in our recent publication (Taguchi et al., 2013), which uses homogenization by passing through needles and sucrose step-gradient. Subcellular fractionation is a very useful technique but usually a
An effective strategy for ligand-mediated pulldown of the HER2/HER3/NRG1I? heterocomplex and cryo-EM structure determination at low sample concentrations.
Trenker, et al.
Methods Enzymol, 667, 633-662 (2023)
Karla Lucía F Alvarez et al.
Veterinary research, 51(1), 123-123 (2020-09-27)
Infectious coryza (IC), an upper respiratory tract disease affecting chickens, is caused by Avibacterium paragallinarum. The clinical manifestations of IC include nasal discharge, facial swelling, and lacrimation. This acute disease results in high morbidity and low mortality, while the course
Ramón Cervantes-Rivera et al.
Bio-protocol, 10(18), e3757-e3757 (2021-03-05)
Gene transcription in bacteria often starts some nucleotides upstream of the start codon. Identifying the specific Transcriptional Start Site (TSS) is essential for genetic manipulation, as in many cases upstream of the start codon there are sequence elements that are
Janine E Melsen et al.
Bio-protocol, 10(23), e3845-e3845 (2021-03-05)
Natural killer (NK) cells are innate immune cells, characterized by their cytotoxic capacity, and chemokine and cytokine secretion upon activation. Human NK cells are identified by CD56 expression. Circulating NK cells can be further subdivided into the CD56bright (~10%) and
Dian Jing et al.
Cell research, 28(8), 803-818 (2018-05-31)
Tissue clearing technique enables visualization of opaque organs and tissues in 3-dimensions (3-D) by turning tissue transparent. Current tissue clearing methods are restricted by limited types of tissues that can be cleared with each individual protocol, which inevitably led to
Saskia V Vijver et al.
Frontiers in immunology, 12, 733561-733561 (2021-10-26)
The tumor microenvironment (TME) is a complex structure comprised of tumor, immune and stromal cells, vasculature, and extracellular matrix (ECM). During tumor development, ECM homeostasis is dysregulated. Collagen remodeling by matrix metalloproteinases (MMPs) generates specific collagen fragments, that can be
Liliana Anjos et al.
MethodsX, 9, 101637-101637 (2022-03-05)
Microalgae have high potential as a resource for sustainable and green protein for food or bioactive molecules. Nonetheless, despite the high protein content of microalgae (40 - 70% dry weight) progress in the characterization of their protein composition remains challenging.
Anne A M J Becker et al.
BMC microbiology, 14, 43-43 (2014-02-20)
Imbalanced feeding regimes may initiate gastrointestinal and metabolic diseases in endangered felids kept in captivity such as cheetahs. Given the crucial role of the host's intestinal microbiota in feed fermentation and health maintenance, a better understanding of the cheetah's intestinal
Eliseo A Eugenin et al.
Journal of neurochemistry, 151(5), 558-569 (2019-08-06)
Chemical and electrical synapses are the two major communication systems that permit cell-to-cell communication within the nervous system. Although most studies are focused on chemical synapses (glutamate, γ-aminobutyric acid, and other neurotransmitters), clearly both types of synapses interact and cooperate
Lucas Bethge et al.
Current protocols in nucleic acid chemistry, 81(1), e109-e109 (2020-05-22)
The reaction between N-hydroxy succinimide (NHS) ester-activated carboxylic acids and amino-modified nucleic acids is commonly used for the post-synthetic modification of oligonucleotides. Here, we report a two-step variation of the method in which the NHS ester is replaced by the
Global analysis of RNA-binding protein dynamics by comparative and enhanced RNA interactome capture.
Joel I Perez-Perri et al.
Nature protocols, 16(1), 27-60 (2020-11-20)
Interactions between RNA-binding proteins (RBPs) and RNAs are critical to cell biology. However, methods to comprehensively and quantitatively assess these interactions within cells were lacking. RNA interactome capture (RIC) uses in vivo UV crosslinking, oligo(dT) capture, and proteomics to identify
Vincent de Bakker et al.
Nature protocols, 17(2), 252-281 (2022-01-09)
CRISPR interference (CRISPRi) is a powerful tool to link essential and nonessential genes to specific phenotypes and to explore their functions. Here we describe a protocol for CRISPRi screenings to assess genome-wide gene fitness in a single sequencing step (CRISPRi-seq).
H H Lee et al.
Journal of food science and technology, 52(12), 7806-7816 (2015-11-26)
Whole grains consumption promotes health benefits, but demonstrates controversial impacts from phytic acid in meeting requirements of good health. Therefore, this study was aimed to determine the nutrient bioaccessibility and antioxidant properties of rice cultivars named "Adan" or "Bario" and
Yuzuru Taguchi et al.
Bio-protocol, 4(11) (2014-06-05)
Here we introduce a protocol for Triton X-114 extraction which we used in our recently-published paper (Taguchi et al., 2013). It is a versatile method to concentrate or partially purify hydrophobic proteins. The presented protocol is based on the protocol
Neha Tiwari et al.
Cell stem cell, 23(4), 557-571 (2018-10-06)
A broad molecular framework of how neural stem cells are specified toward astrocyte fate during brain development has proven elusive. Here we perform comprehensive and integrated transcriptomic and epigenomic analyses to delineate gene regulatory programs that drive the developmental trajectory
Roberto Alcántara et al.
STAR protocols, 2(4), 100878-100878 (2021-10-05)
Here, we describe a detailed step-by-step protocol to detect SARS-CoV-2 RNA using RT-PCR-mediated amplification and CRISPR/Cas-based visualization. The optimized assay uses basic molecular biology equipment such as conventional thermocyclers and transilluminators for qualitative detection. Alternatively, a fluorescence plate reader can
Jacob P van Vloten et al.
Molecular therapy. Methods & clinical development, 13, 154-166 (2019-02-23)
Immunotherapies are at the forefront of the fight against cancers, and researchers continue to develop and test novel immunotherapeutic modalities. Ideal cancer immunotherapies induce a patient's immune system to kill their own cancer and develop long-lasting immunity. Research has demonstrated
Clara Alice Musi et al.
BMC biology, 19(1), 256-256 (2021-12-17)
Rett syndrome (RTT) is a monogenic X-linked neurodevelopmental disorder characterized by loss-of-function mutations in the MECP2 gene, which lead to structural and functional changes in synapse communication, and impairments of neural activity at the basis of cognitive deficits that progress
Kimiya Memarzadeh et al.
Cancer biology & therapy, 20(12), 1416-1429 (2019-09-03)
Neuroblastoma is the most common malignancy in infants. Overexpression of the epidermal growth factor receptor (EGFR) in neuroblastoma tumors underlies resistance to chemotherapeutics. UBE4B, an E3/E4 ubiquitin ligase involved in EGFR degradation, is located on chromosome 1p36, a region in
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