Forward genetic screens to identify circadian rhythm mutants in mice.

abstract：:Nitric oxide formation in ischemia-reperfusion injury can be identified and measured directly using EPR of nitroso-heme complexes comprising NO bound to either Mb or Hb-Fe2+. This article described the successful use of this method to detect and quantify the generation of NO formed independently of nitric oxide syntha...

journal_title：Methods in enzymology

authors： Lepore DA

更新日期：2002-01-01 00:00:00

abstract：:Given the increasing evidence of oxidative stress in AD brain and studies from different perspectives that appear to show a converging, central role for A beta in the pathogenesis and etiology of AD, insight into A beta-associated free radical oxidative stress will likely lead to a greater understanding of AD and, pot...

journal_title：Methods in enzymology

authors： Butterfield DA,Yatin SM,Varadarajan S,Koppal T

更新日期：1999-01-01 00:00:00

abstract：:IF-liposomes are formed by a unique process that involves fusing small liposomes into interdigitated lipid sheets, using either ethanol or hydrostatic pressure. The interdigitation-fusion method requires liposome formulations with lipids that form the L beta I phase. Preparing ethanol-induced IF-liposomes is simple an...

journal_title：Methods in enzymology

authors： Ahl PL,Perkins WR

更新日期：2003-01-01 00:00:00

abstract：:Essentially the same steps are required to solve the crystal structure of a serpin as for any other protein: produce and purify protein, grow crystals, collect diffraction data, find estimates of the phase angles, and then refine and validate the structure. For the phasing step, experimental phasing methods involving ...

journal_title：Methods in enzymology

authors： Read RJ,Zhou A,Stein PE

更新日期：2011-01-01 00:00:00

abstract：:A synthetic ligand regulable system for gene transfer and expression has been developed in our laboratory based on mechanistic studies of steriod hormone receptor and transcriptional regulation. This gene switch system possesses most of the important features that are required for application of the system in biologic...

journal_title：Methods in enzymology

authors： Ye X,Schillinger K,Burcin MM,Tsai SY,O'Malley BW

更新日期：2002-01-01 00:00:00

abstract：:In vivo genome manipulation through site-directed mutagenesis and chromosome rearrangements has been hindered by the difficulty in achieving high frequencies of targeting and the intensive labor required to create altered genomes that do not contain any heterologous sequence. Here we describe our approach, referred to...

journal_title：Methods in enzymology

authors： Storici F,Resnick MA

更新日期：2006-01-01 00:00:00

abstract：:Lymphocytes proliferate in response to several stimuli. In many situations, a rapid lymphocyte expansion, or the identification of a slow dividing cell subpopulation may be of great interest. Thus, it is necessary to perform reliable assays to study and compare lymphocyte subsets proliferation. For this purpose, carbo...

journal_title：Methods in enzymology

authors： Terrén I,Orrantia A,Vitallé J,Zenarruzabeitia O,Borrego F

更新日期：2020-01-01 00:00:00

abstract：:Antibodies will be immobilized on a cyanogen bromide-activated Sepharose for subsequent use in pull-down assays or immunoaffinity purification. ...

journal_title：Methods in enzymology

authors： Kavran JM,Leahy DJ

更新日期：2014-01-01 00:00:00

abstract：:Galectins are a family of animal lectins with affinity for beta-galactosides. By using recombinant proteins, a number of galectins have been shown to interact with cell-surface and extracellular matrix glycoconjugates through lectin-carbohydrate interactions. Through this action, they can affect a variety of cellular ...

journal_title：Methods in enzymology

authors： Hsu DK,Yang RY,Liu FT

更新日期：2006-01-01 00:00:00

abstract：:Primary mixed liver cells were isolated from rats that had been fed an amino acid (AA) diet in which natural protein was replaced with a defined mixture of pure AAs. Nitric oxide (NO) production from these cells in vitro was monitored using a nitric oxide (NO)-selective fluorescent probe, diaminofluorescein, followed ...

journal_title：Methods in enzymology

authors： Kotake Y,Kishida H,Nakae D,Floyd RA

更新日期：2005-01-01 00:00:00

abstract：:The growth and survival of cancer cells are often driven by constitutive activity in the mitogen-activated protein kinase (MAPK) and phospho-inositide 3-kinase (PI3K)/AKT signaling pathways. Activity in these signal transduction cascades is known to contribute to the uncontrolled growth and resistance to apoptosis tha...

journal_title：Methods in enzymology

authors： Paraiso KH,Van Der Kooi K,Messina JL,Smalley KS

更新日期：2010-01-01 00:00:00

abstract：:Enzyme immobilization is a widespread empiric technology to achieve more stable, active and reusable enzymes. The empiricism can be reduced by the application of rational design procedures employing bioinformatic tools, engineered-proteins and detailed analyses of existent data. In this chapter, we describe relevant a...

journal_title：Methods in enzymology

authors： Sastre DE,Reis EA,Marques Netto CGC

更新日期：2020-01-01 00:00:00

abstract：:Three-dimensional (3D) cryoelectron microscopy reconstruction methods are uniquely able to reveal structures of many important macromolecules and macromolecular complexes. EMDataBank.org, a joint effort of the Protein Databank in Europe (PDBe), the Research Collaboratory for Structural Bioinformatics (RCSB), and the N...

journal_title：Methods in enzymology

authors： Lawson CL

更新日期：2010-01-01 00:00:00

abstract：:Mdm2 is a negative regulator of p53 activity and functions as an E3 ubiquitin ligase of p53. Inhibition of mdm2 E3 ligase activity will block ubiquitination and subsequent proteasome-mediated degradation of p53, resulting in the stabilization of p53 protein that could lead to the restoration of its tumor-suppressor ac...

journal_title：Methods in enzymology

authors： Auger KR,Copeland RA,Lai Z

更新日期：2005-01-01 00:00:00

abstract：:The circadian clock exists to synchronize inner physiology with the external world, allowing life to anticipate and adapt to the continual changes that occur in an organism's environment. The clock architecture is highly conserved, present in almost all major branches of life. Within eukaryotes, the filamentous fungus...

journal_title：Methods in enzymology

authors： Hurley J,Loros JJ,Dunlap JC

更新日期：2015-01-01 00:00:00

abstract：:We describe here the methods we have used to generate selective peptide inhibitors and activators of PKC-mediated signaling. These approaches should be applicable to any signaling event that is dependent on protein-protein interaction. Furthermore, targeting downstream enzymes in signal transduction has been notorious...

journal_title：Methods in enzymology

authors： Schechtman D,Mochly-Rosen D

更新日期：2002-01-01 00:00:00

abstract：:Many methods have been developed for the cloning of PCR products. These methods include blunt-end cloning, TA cloning, and using restriction sites incorporated into the PCR primers. The restrictionless cloning technique allows efficient directional cloning of PCR products into any cloning site within a vector regardle...

journal_title：Methods in enzymology

authors： Algire MA

更新日期：2013-01-01 00:00:00

abstract：:The reverse dot-blot method is a simple and rapid diagnostic procedure that allows screening of sample for a variety of mutations/polymorphisms in a single hybridization reaction. Several methods of immobilizing the oligonucleotide probes are discussed. The reverse dot-blot method has several unique properties that ar...

journal_title：Methods in enzymology

authors： Kawasaki E,Saiki R,Erlich H

更新日期：1993-01-01 00:00:00

abstract：:This chapter provides the methodologies employed to study the polymorphism of human apoE. These and other related studies have advanced our understanding of the structure and function of this protein as follows: The complex array of human apoE observed by two-dimensional gel electrophoresis results from genetic variat...

journal_title：Methods in enzymology

authors： Zannis VI

更新日期：1986-01-01 00:00:00

abstract：:A significant proportion of RNA decay in eukaryotes takes place in the nucleus. In addition to targeting nongenic RNAs such as products of processing and spurious transcription events, nuclear RNA decay also removes aberrant transcripts from regular genes. Moreover, a small list of genes utilizes nuclear RNA turnover ...

journal_title：Methods in enzymology

authors： Schmid M,Rougemaille M,Libri D,Jensen TH

更新日期：2008-01-01 00:00:00

abstract：:Recent studies using sulfotransferase-deficient mice have revealed various physiological functions of sulfated glycans. Studies using gene-targeted mice deficient in both N-acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST)-1 and GlcNAc6ST-2 showed that these sulfotransferases play critical roles in lymphocyte homing....

journal_title：Methods in enzymology

authors： Kawashima H

更新日期：2010-01-01 00:00:00

abstract：:Enzymology is approaching an era where many problems can benefit from computational studies. While ample challenges remain in quantitatively predicting behavior for many enzyme systems, the insights that often come from computations are an important asset for the enzymology community. Here we provide a primer for enzy...

journal_title：Methods in enzymology

authors： Roston D,Cui Q

更新日期：2016-01-01 00:00:00

abstract：:In vitro assembly of eukaryotic translation initiation complexes requires purification of ribosomal subunits, eukaryotic initiation factors, and initiator tRNA from natural sources and therefore yields only limited material for functional and structural studies. In this chapter, we describe a robust, affinity chromato...

journal_title：Methods in enzymology

authors： Locker N,Lukavsky PJ

更新日期：2007-01-01 00:00:00

abstract：:RNA helicases are associated with every aspect of RNA metabolism and function. A diverse range of RNA helicases are encoded by essentially every organism. While RNA helicases alter gene expression, RNA helicase expression is itself regulated, frequently in response to abiotic stress. Photosynthetic cyanobacteria prese...

journal_title：Methods in enzymology

authors： Owttrim GW

更新日期：2012-01-01 00:00:00

abstract：:In mice with lupus nephritis qualitative changes in anti-DNA antibodies occur, such as IgG switch and increased cationic charge, to render these antibodies pathogenic. Pathogenic anti-DNA idiotypes can be encoded by genes of a normal mouse strain such as SWR, where they remain dormant. When the normal mice are crossed...

journal_title：Methods in enzymology

authors： Datta SK

更新日期：1988-01-01 00:00:00

abstract：:Histone variants and posttranslational modifications (PTMs) are essential for epigenetic regulation of transcriptional expression. Single and/or combinatorial PTMs of histones play important roles in development and disease formation. Mass spectrometry (MS) has been a powerful tool to study histone variants and PTMs, ...

journal_title：Methods in enzymology

authors： Lin S,Garcia BA

更新日期：2012-01-01 00:00:00

abstract：:The epitope tagging approach offers advantages of economy, universality, and precision over the use of antibodies raised directly against a protein of interest. The latter strategy promises a potentially greater diversity of reagents and obviates the need to modify the protein, but it may not yield sufficiently high-a...

journal_title：Methods in enzymology

authors： Kolodziej PA,Young RA

更新日期：1991-01-01 00:00:00

abstract：:A simple and convenient method is described to determine primary deuterium kinetic isotope effects (1°DKIEs) on reactions where the hydron incorporated into the reaction product is derived from solvent water. The 1°DKIE may be obtained by 1H NMR analyses as the ratio of the yields of H- and D-labeled products from a r...

journal_title：Methods in enzymology

authors： Amyes TL,Richard JP

更新日期：2017-01-01 00:00:00

abstract：:The kinetics of biomolecular systems can be quantified by calculating the stochastic rate constants that govern the biomolecular state vs time trajectories (i.e., state trajectories) of individual biomolecules. To do so, the experimental signal vs time trajectories (i.e., signal trajectories) obtained from observing i...

journal_title：Methods in enzymology

authors： Kinz-Thompson CD,Bailey NA,Gonzalez RL Jr

更新日期：2016-01-01 00:00:00

abstract：:Survival of macrophages, which serve as the first-line defense against invading pathogens by invoking the overproduction of highly toxic peroxynitrite (ONOO-), depends on their ability to maintain the intracellular GSH level and to induce the expression of heme oxygenase-1 (HO-1). The ONOO- is produced by macrophages ...

journal_title：Methods in enzymology

authors： Srisook K,Kim C,Cha YN

更新日期：2005-01-01 00:00:00