Highly Modular Bioluminescent Sensors for Small Molecules and Proteins.

abstract：:This chapter described the preparation and fractionation of libraries of M13 phage displaying proteins as fusions to the major coat protein. High titer (10(13) pfu/ml) phage libraries can readily be generated using a single vector and the level of display surpasses that of gene III fusion phage. Since the synthetic VI...

journal_title：Methods in enzymology

authors： Roberts BL,Markland W,Ladner RC

更新日期：1996-01-01 00:00:00

abstract：:White adipose tissue (WAT) is a heterogeneous tissue composed of lipid-filled adipocytes and several nonadipocyte cell populations, including endothelial, blood, uncharacterized stromal, and adipocyte precursor cells. Although lipid-filled adipocytes account for the majority of WAT volume and mass, nonadipocyte cell p...

journal_title：Methods in enzymology

authors： Church CD,Berry R,Rodeheffer MS

更新日期：2014-01-01 00:00:00

abstract：:Growing appreciation for the biogeochemical significance of uncultured microorganisms is changing the focus of environmental microbiology. Techniques designed to investigate microbial metabolism in situ are increasingly popular, from mRNA-targeted fluorescence in situ hybridization (FISH) to the "-omics" revolution, i...

journal_title：Methods in enzymology

authors： Dekas AE,Orphan VJ

更新日期：2011-01-01 00:00:00

abstract：:Nonsense-mediated mRNA decay (NMD) is an evolutionarily conserved mechanism of specific degradation of transcripts with a premature stop codon. NMD eliminates aberrant mRNAs arising from mutations, alternative splicing, and other events in cells. In addition, many normal transcripts undergo NMD. Recent studies demonst...

journal_title：Methods in enzymology

authors： Gurskaya NG,Pereverzev AP,Staroverov DB,Markina NM,Lukyanov KA

更新日期：2016-01-01 00:00:00

abstract：:Anomalous small angle X-ray scattering (ASAXS) exploits contrast variation methods to highlight the scattering from one elemental component in a multielement sample, such as one ion species in an ion-DNA system. The ASAXS method has been applied to measure ions condensed around short nucleic acid duplexes. This chapte...

journal_title：Methods in enzymology

authors： Pabit SA,Finkelstein KD,Pollack L

更新日期：2009-01-01 00:00:00

abstract：:Approximately one-third of all newly translated polypeptides interact with the endoplasmic reticulum (ER), an event that is essential to target these nascent proteins to distinct compartments within the cell or to the extracellular milieu. Thus, the ER houses molecular chaperones that augment the folding of this diver...

journal_title：Methods in enzymology

authors： Brodsky JL

更新日期：2010-01-01 00:00:00

abstract：:Neurons are probably the most highly polarized cell type and typically develop a single axon and several dendrites. The establishment of a polarized morphology and the functional specialization of axonal and dendritic compartments are essential steps in the differentiation of neurons. Primary cultures of dissociated h...

journal_title：Methods in enzymology

authors： Schwamborn JC,Li Y,Püschel AW

更新日期：2006-01-01 00:00:00

abstract：:Autophagy is a catabolic process that is crucial for cellular homeostasis and adaptive response to changing environments. Importantly, autophagy has been shown to be induced in many longevity-associated scenarios and to be required to maintain lifespan extension. Notably, autophagy is a highly conserved cellular proce...

journal_title：Methods in enzymology

authors： Kainz K,Tadic J,Zimmermann A,Pendl T,Carmona-Gutierrez D,Ruckenstuhl C,Eisenberg T,Madeo F

更新日期：2017-01-01 00:00:00

abstract：:Cell-free (CF) protein expression has emerged as one of the most efficient production platforms for membrane proteins. Central bottlenecks prevalent in conventional cell-based expression systems such as mistargeting, inclusion body formation, degradation as well as product toxicity can be addressed by taking advantage...

journal_title：Methods in enzymology

authors： Henrich E,Dötsch V,Bernhard F

更新日期：2015-01-01 00:00:00

abstract：:Nonsense-mediated mRNA decay (NMD) is activated by exon-junction complexes (EJCs) that are located downstream of the termination codon of the substrate mRNAs. This situation can be imitated by tethering components of the EJC to the 3' untranslated region (3' UTR) of a reporter mRNA. Here we describe the detailed use o...

journal_title：Methods in enzymology

authors： Gehring NH,Hentze MW,Kulozik AE

更新日期：2008-01-01 00:00:00

abstract：:This protocol describes Coomassie brilliant blue staining, one of the most common methods of detecting proteins in polyacrylamide gels (PAGE). ...

journal_title：Methods in enzymology

authors： Brunelle JL,Green R

更新日期：2014-01-01 00:00:00

abstract：:Powerful specialized software is essential for managing, quantifying, and ultimately deriving scientific insight from results of a microarray experiment. We have developed a suite of software applications, known as TM4, to support such gene expression studies. The suite consists of open-source tools for data managemen...

journal_title：Methods in enzymology

authors： Saeed AI,Bhagabati NK,Braisted JC,Liang W,Sharov V,Howe EA,Li J,Thiagarajan M,White JA,Quackenbush J

更新日期：2006-01-01 00:00:00

abstract：:Numerous aspects of the immune system operate on the basis of complex regulatory networks that are amenable to mathematical and computational modeling. Several modeling frameworks have recently been applied to simulating the immune system, including systems of ordinary differential equations, delay differential equati...

journal_title：Methods in enzymology

authors： Kim PS,Levy D,Lee PP

更新日期：2009-01-01 00:00:00

abstract：:The methodologies described in this chapter allow the reproducible preparation of native high-molecular-weight synthetase complexes of leuRL. These complexes have the ability to preferentially utilize extracellular leucine immediately upon transport and are likely the forms of the enzyme most important in the utilizat...

journal_title：Methods in enzymology

authors： Hampel A,Tritz R

更新日期：1988-01-01 00:00:00

abstract：:The current treatment of neurological and psychiatric diseases is far beyond being satisfactory. In addition to highly complex disease mechanisms, the blood-brain barrier (BBB) also remains as a challenge by limiting the delivery of the majority of currently available therapeutics to the central nervous system. Severa...

journal_title：Methods in enzymology

authors： Yemişci M,Gürsoy-Özdemir Y,Caban S,Bodur E,Capan Y,Dalkara T

更新日期：2012-01-01 00:00:00

abstract：:Two-hybrid systems are one of the most popular, preferred, cost effective, and scalable in vivo genetic approaches for screening protein-protein interactions. A number of variants of yeast and bacterial two-hybrid systems exist, rendering them ideal for modern, flexible proteomics-driven studies. For mapping protein i...

journal_title：Methods in enzymology

authors： Mehla J,Caufield JH,Sakhawalkar N,Uetz P

更新日期：2017-01-01 00:00:00

abstract：:Current methodologies available to quantify changes in mitochondrial turnover are limited to pulse-chase assays or specific assays that quantify mitophagy. Accordingly, new tools that can assess mitochondrial turnover are needed for the study of cellular, subcellular, and spatial parameters of mitochondrial turnover a...

journal_title：Methods in enzymology

authors： Trudeau KM,Gottlieb RA,Shirihai OS

更新日期：2014-01-01 00:00:00

abstract：:There is a great need for practicing scientists to volunteer their time and expertise in the K-12th grade science classroom. We have found that bioluminescence is a fun and exciting way to teach basic science concepts and is an excellent tool for the volunteering scientist. We have had very positive reactions from bot...

journal_title：Methods in enzymology

authors： Hammer M,Andrade JD

更新日期：2000-01-01 00:00:00

abstract：:In polarized epithelial cells, the tight junction has been ascribed several functions including the regulation of the paracellular permeability, an impediment to the diffusion of molecules between the apical and basolateral domains, a site of delivery of transport vesicles for basolateral proteins, and a scaffold for ...

journal_title：Methods in enzymology

authors： Luton F

更新日期：2005-01-01 00:00:00

abstract：:The combined use of rotating-frame relaxation methods, temperature-dependent measurements of line shapes and magnetization transfer experiments allows in favorable cases the examination in some detail of exchange processes that occur on the millisecond time scale. It is possible to determine not only the rate constant...

journal_title：Methods in enzymology

authors： Lane AN,Lefèvre JF

更新日期：1994-01-01 00:00:00

abstract：:Reliable detection of keratins in tissues is important for investigating their physiological role and for using keratin expression as a biomarker in medical diagnostics. A particular challenge for the detection of keratins by immunofluorescence microscopy or immunohistochemistry relates to the fact that keratin interm...

journal_title：Methods in enzymology

authors： Stumptner C,Gogg-Kamerer M,Viertler C,Denk H,Zatloukal K

更新日期：2016-01-01 00:00:00

abstract：:Benzylisoquinoline alkaloids (BIAs) constitute a diverse class of plant secondary metabolites that includes the opiate analgesics morphine and codeine. Collectively, BIAs exhibit a myriad of pharmacological activities, including antimicrobial, antitussive, antispasmodic, and anticancer properties. Despite 2500 known B...

journal_title：Methods in enzymology

authors： Pyne ME,Narcross L,Fossati E,Bourgeois L,Burton E,Gold ND,Martin VJ

更新日期：2016-01-01 00:00:00

abstract：:Structure-based protein design is a powerful technique with great potential. Challenges in two areas limit performance: structure scoring and sequence-structure searching. Many of the functions used to describe the relationship between protein sequence and energy are computationally expensive to evaluate, and the spac...

journal_title：Methods in enzymology

authors： Negron C,Keating AE

更新日期：2013-01-01 00:00:00

abstract：:Using the expression strategies described here, we have demonstrated a model system whereby the sequential signaling events involved in cell proliferation and subsequent transformation regulated by G alpha 12 can be investigated. The model system presented here can also be used to study the temporal interrelationships...

journal_title：Methods in enzymology

authors： Dermott JM,Dhanasekaran N

更新日期：2002-01-01 00:00:00

abstract：:The unexpected discovery of graphene and especially the follow-up explosion of interest in its properties and applications marked the beginning of a new carbon era. Graphene-based nanostructured materials are highly useful because they show great promise in the field of biotechnology and biomedicine. Owing to their un...

journal_title：Methods in enzymology

authors： Khan M,Husain Q,Asmat S

更新日期：2018-01-01 00:00:00

abstract：:Rarely is any solution simply solute and water. In vivo, solutes, such as proteins and nucleic acids, swim in a sea of water, salts, ions, small molecules, and lipids, not to mention other macromolecules. In vitro, virtually all solutions contain a mixture of aqueous solvents, or "cosolvents" [i.e., solvent(s) in addi...

journal_title：Methods in enzymology

authors： Beck DA,Bennion BJ,Alonso DO,Daggett V

更新日期：2007-01-01 00:00:00

abstract：:Peptides corresponding to the calmodulin-binding domain of MLCK and other target enzymes promise to be useful tools in the study of calmodulin action. Not only will they help to elucidate the molecular interactions of calmodulin with specific target enzymes, but they should also prove to be useful calmodulin antagonis...

journal_title：Methods in enzymology

authors： Blumenthal DK,Krebs EG

更新日期：1987-01-01 00:00:00

abstract：:The anaphase-promoting complex (APC) is a central regulator of the eukaryotic cell cycle and functions as an E3 ubiquitin protein ligase to catalyze the ubiquitination of a number of cell cycle regulatory proteins. The APC contains at least 13 subunits in addition to two activator subunits, Cdc20 and Cdh1, that associ...

journal_title：Methods in enzymology

authors： Passmore LA,Barford D,Harper JW

更新日期：2005-01-01 00:00:00

abstract：:This article describes the methods and techniques used to produce mutagenized mice to conduct high-throughput forward genetic screens for circadian rhythm mutants in the mouse. In particular, we outline methods to safely prepare and administer the chemical mutagen N-nitroso-N-ethylurea (ENU) to mice. We also discuss t...

journal_title：Methods in enzymology

authors： Siepka SM,Takahashi JS

更新日期：2005-01-01 00:00:00

abstract：:Formation of 5-methyluridine (ribothymidine) at position 54 of the T-psi loop of tRNA is catalyzed by site-specific tRNA methyltransferases (tRNA[uracil-54,C5]-MTases). In eukaryotes and many bacteria, the methyl donor for this reaction is generally S-adenosyl-L-methionine (S-AdoMet). However, in other bacteria, like ...

journal_title：Methods in enzymology

authors： Urbonavicius J,Brochier-Armanet C,Skouloubris S,Myllykallio H,Grosjean H

更新日期：2007-01-01 00:00:00