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Methods for Biophysical Characterization of SznF, a Member of the Heme-Oxygenase-Like Diiron Oxidase/Oxygenase Superfamily
Nonheme diiron enzymes harness the chemical potential of oxygen to catalyze challenging reactions in biology. In their resting state, these enzymes...
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Dissecting Selectivity Determinants of Small-Molecule Inhibitors of SH2 Domains Via Fluorescence Polarization Assays
Fluorescence polarization (FP) assays can be used to identify small-molecule inhibitors that bind to SH2 domain-containing proteins. We have...
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A simple and effective method to remove pigments from heterologous secretory proteins expressed in Pichia pastoris
Pichia pastoris is a popular yeast host for high-level heterologous expression of proteins on an industrial scale owing to its reliable expression,...
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Purification, Reconstitution, and Functional Analysis of Connexin Hemichannels
Connexins are the proteins that form the gap junction channels that are essential for cell-to-cell communication. These channels are formed by...
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Bacterial Two Component Systems: Overexpression and Purification: In Vitro and In Vivo Inhibitor Screens
Bacterial histidine kinases are promising targets for new antimicrobial agents. In antibacterial therapy, such agents could inhibit bacterial growth...
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Quantification of Chromogranin A and Its Fragments in Biological Fluids
Human chromogranin A (CgA), a 439-residue long neurosecretory protein, can serve as a circulating biomarker for a wide range of neuroendocrine...
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Remediation of dairy wastewater by Euglena gracilis WZSL mutant and β-glucan production
According to the concept of circular economy, food industry wastewater should be considered a secondary raw material that could be exploited, thus...
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Reconstitution of Mammalian Mitochondrial Translation System Capable of Long Polypeptide Synthesis
Mammalian mitochondria have their own dedicated protein synthesis system, which produces 13 essential subunits of the oxidative phosphorylation...
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A Method for Urinary Trypsin Inhibitor (UTI) Purification Combining Anion-Exchange Chromatography Enrichment and Preparative SDS-PAGE
This chapter describes a method for the purification of urinary trypsin inhibitor (UTI), a small chondroitin sulfate proteoglycan with Ser-proteinase...
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Observing isolated synaptic vesicle association and fusion ex vivo
Here, we present a protocol for isolating functionally intact glutamatergic synaptic vesicles from whole-mouse brain tissue and using them in a...
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Isothermal Titration Calorimetry for Fragment-Based Analysis of Ion Channel Interactions
Ion channels are membrane proteins that may also have intracellular and extracellular domains that interact with other ligands. In many cases, these...
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The root growth reduction in response to mechanical stress involves ethylene-mediated microtubule reorganization and transmembrane receptor-mediated signal transduction in Arabidopsis
Key messageWe found that mutations in a Ca 2+ -permeable mechanosensitive channel MCA1, an ethylene-regulated microtubule-associated protein WDL5,...
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Extracellular Vesicles: The Challenges on the Way and Engineering Perspectives
Extracellular vesicles (EVs) are found in every body fluid and are known to be released by every human cell. So far, EVs have been classified into...
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Detecting Poly (ADP-Ribose) In Vitro and in Cells Using PAR Trackers
ADP-ribosylation (ADPRylationADP-ribosylation (ADPRylation)) is a reversible posttranslational modification resulting in the covalent attachment of...
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Imaging Chromatin Accessibility by Assay of Transposase-Accessible Chromatin with Visualization
Chromatin accessibility is one of the fundamental structures regulating genome functions including transcription and DNA repair. Recent technological...
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Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology
According to the free hormone hypothesis, biological activity of a certain hormone is best reflected by free rather than total hormone...
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Effect of preservation fluid contamination and associated possible donor-derived infections on early postoperative prognosis in kidney transplant recipients
BackgroundThe study aims to analyze the epidemiology of preservation fluid (PF) contamination and investigate the impact of PF contamination and...
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A simple and unified protocol to purify all seven Escherichia coli RNA polymerase sigma factors
RNA polymerase sigma factors are indispensable in the process of bacterial transcription. They are responsible for a given gene’s promoter region...
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ATAC-See: A Tn5 Transposase-Mediated Assay for Detection of Chromatin Accessibility with Imaging
Assay of transposase-accessible chromatin with visualization (ATAC-see), a transposase-mediated imaging technology that enables direct imaging of the...
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Deficiency in PHD2-mediated hydroxylation of HIF2α underlies Pacak-Zhuang syndrome
Pacak-Zhuang syndrome is caused by mutations in the EPAS1 gene, which encodes for one of the three hypoxia-inducible factor alpha (HIFα) paralogs...
