Quantitative analysis of microtubule self-assembly kinetics and tip structure.

abstract：:Gas chromatography-mass spectrometry (GC-MS) is one of the three most popular analytical platforms for metabolomics and is largely employed for the study of oncometabolism. Large volumes of data are usually generated in a GC-MS experiment, and many analytical steps are required to extract biologically relevant informa...

journal_title：Methods in enzymology

authors： Koo I,Wei X,Zhang X

更新日期：2014-01-01 00:00:00

abstract：:Hydrogen sulfide is an endogenous gaseous mediator that plays important roles in many physiological processes in microbes, plants, and animals. This chapter focuses on the important roles of hydrogen sulfide in protecting tissues against injury, promoting the repair of damage, and downregulating the inflammatory respo...

journal_title：Methods in enzymology

authors： Gemici B,Wallace JL

更新日期：2015-01-01 00:00:00

abstract：:Cholestatic syndromes are inborn or acquired disorders of bile formation. In recent years, several inherited cholestatic syndromes were characterized at the molecular level: progressive familial intrahepatic cholestasis (PFIC) and benign recurrent intrahepatic cholestasis (BRIC). Both PFIC and BRIC were divided phenot...

journal_title：Methods in enzymology

authors： Kubitz R,Keitel V,Häussinger D

更新日期：2005-01-01 00:00:00

abstract：:Chaperone-mediated autophagy (CMA) is a selective type of autophagy responsible for the lysosomal degradation of soluble cytosolic proteins. In contrast to other forms of autophagy where cargo is sequestered and delivered to lysosomes through membrane fusion/excision, CMA substrates reach the lysosomal lumen after dir...

journal_title：Methods in enzymology

authors： Kaushik S,Cuervo AM

更新日期：2009-01-01 00:00:00

abstract：:Complex synthetic schemes catalyzed by multienzyme systems immobilized on solid materials are gaining momentum in chemical biomanufacturing. These systems harness the high chemo-, regio-, and stereoselectivity of the enzymes and the recyclability of the heterogeneous catalysts. Moreover, when the enzymes become part o...

journal_title：Methods in enzymology

authors： López-Gallego F

更新日期：2019-01-01 00:00:00

abstract：:Immunogenic cell death (ICD), induced by certain anticancer chemotherapeutics, leads to the emission of danger associated molecular patterns (DAMP) by cancer cells, which facilitates the attraction, activation and maturation of dendritic cells (DC) as well as the subsequent priming of effector T cells. In this context...

journal_title：Methods in enzymology

authors： Cerrato G,Liu P,Martins I,Kepp O,Kroemer G

更新日期：2020-01-01 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：:Small GTPases of the Ras and Rho families are among the most studied signaling proteins and represent promising therapeutic targets for human neoplastic disease. Despite the high level of interest in these proteins, direct analysis of most aspects of Ras protein biology in living cells has not been possible, because m...

journal_title：Methods in enzymology

authors： Westwick JK,Michnick SW

更新日期：2006-01-01 00:00:00

abstract：:T lymphocytes express on their surface a heterodimeric αβ receptor, called the T cell receptor (TCR), which recognizes foreign antigens. Unlike antibodies, the recognition requires both an antigenic peptide epitope and a protein encoded by the major histocompatibility complex (MHC). In contrast to conventional antibod...

journal_title：Methods in enzymology

authors： Stone JD,Chervin AS,Aggen DH,Kranz DM

更新日期：2012-01-01 00:00:00

abstract：:Biotinylated molecules are extensively employed in bioanalytics and biotechnology. The currently available assays for quantification of biotin groups suffer from low sensitivity, low accuracy, or provide highly variable responses for different biotin derivatives. We developed a competitive binding assay in which avidi...

journal_title：Methods in enzymology

authors： Oberbichler E,Wiesauer M,Schlögl E,Stangl J,Faschinger F,Knör G,Gruber HJ,Hytönen VP

更新日期：2020-01-01 00:00:00

abstract：:Cancer has become the leading cause of death in the developed world and has remained one of the most difficult diseases to treat. One of the difficulties in treating cancer is that conventional chemotherapies often have unacceptable toxicities toward normal cells at the doses required to kill tumor cells. Thus, the de...

journal_title：Methods in enzymology

authors： Cantor JR,Panayiotou V,Agnello G,Georgiou G,Stone EM

更新日期：2012-01-01 00:00:00

abstract：:The methods described here concern the use specialized glassware to achieve accurate sub-micromolar to low-millimolar dioxygen concentrations specifically for the study of flavoproteins. Since the early 1960s, these methods have been passed through the lineage of flavin enzymologists almost exclusively as an oral trad...

journal_title：Methods in enzymology

authors： Moran GR

更新日期：2019-01-01 00:00:00

abstract：:In principle, the long emission lifetimes of lanthanide chelates should enable their ultrasensitive detection in biological systems by time-resolved optical microscopy. However, most lanthanide-imaging systems cannot achieve sensitivities that exceed those of conventional fluorescence microscopes, since they are limit...

journal_title：Methods in enzymology

authors： Ciepla P,Cho U,Chen JK

更新日期：2020-01-01 00:00:00

abstract：:Monte Carlo simulations of neighbor exclusion models have been used to demonstrate the importance of collecting and fitting data over a wide range of saturation. Low saturation data are important for good estimates of the affinity K of a drug or protein for the lattice site. High saturation data are important for dist...

journal_title：Methods in enzymology

authors： Correia JJ,Chaires JB

更新日期：1994-01-01 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：:In this article, we have described methods used to purify Rnt1p and study its biochemical properties. Rnt1p can be easily purified from bacteria as N-terminal His6-tagged protein and its activity may be monitored in vitro. Rnt1p cleaves the RNA by binding to a cleavage site followed by hydrolysis and product release. ...

journal_title：Methods in enzymology

authors： Lamontagne B,Elela SA

更新日期：2001-01-01 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：:Phospholipase A(2)s (PLA(2)s) are a superfamily of enzymes involved in production of a wide variety of lipid mediators, including arachidonic acid, lysophospholipids, platelet activation factor, and eicosanoids. Fluorescence-based, real-time cellular activity assays for PLA(2)s have been developed as a tool for studyi...

journal_title：Methods in enzymology

authors： Manna D,Cho W

更新日期：2007-01-01 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：:Intraflagellar transport (IFT) involves the movement of large proteinaceous particles or trains along the length of ciliary and flagellar axonemal microtubules. The particles contain multiple copies of two protein complexes. As isolated from the flagellated model organism, Chlamydomonas reinhardtii, IFT A contains 6 d...

journal_title：Methods in enzymology

authors： Behal RH,Cole DG

更新日期：2013-01-01 00:00:00

abstract：:Several genetically encoded fluorescent biosensors for Ras family GTPases have been developed that permit spatiotemporal analysis of the activation of these signaling molecules in living cells. We describe here the use of the simplest of these probes, the Ras binding domain (RBD) of selected effectors fused with green...

journal_title：Methods in enzymology

authors： Bivona TG,Quatela S,Philips MR

更新日期：2006-01-01 00:00:00

abstract：:Reference standard materials (RSMs) exist for a variety of biologics including vaccines but are not readily available for gene therapy vectors. To date, a recombinant adeno-associated virus serotype 2 RSM (rAAV2 RSM) has been produced and characterized and was made available to the scientific community in 2010. In add...

journal_title：Methods in enzymology

authors： Moullier P,Snyder RO

更新日期：2012-01-01 00:00:00

abstract：:RNA and DNA carry out diverse functions in biology including catalysis, splicing, gene regulation, and storage of genetic information. Interest has grown in understanding how nucleic acids perform such sophisticated functions given their limited molecular repertoire. RNA can fold into diverse shapes that often perturb...

journal_title：Methods in enzymology

authors： Thaplyal P,Bevilacqua PC

更新日期：2014-01-01 00:00:00

abstract：:Single-molecule Förster resonance energy transfer (smFRET) is a versatile tool for studying biomolecules in a quantitative manner. Multiple conformations within and interactions between biomolecules can be detected and their kinetics can be determined. Thus, smFRET has become an essential tool in enzymology. Ordinary ...

journal_title：Methods in enzymology

authors： Götz M,Wortmann P,Schmid S,Hugel T

更新日期：2016-01-01 00:00:00

abstract：:Mitochondrial uncoupling protein-2 (UCP2) regulates glucose-stimulated insulin secretion (GSIS) by pancreatic beta cells-the physiological role of the beta cell UCP2 remains a subject of debate. Experimental studies informing this debate benefit from reliable measurements of UCP2 protein level and activity. In this ch...

journal_title：Methods in enzymology

authors： Barlow J,Hirschberg V,Brand MD,Affourtit C

更新日期：2013-01-01 00:00:00

abstract：:The programmed death 1 (PD-1) and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) are negative regulators of T-cell immune function. Removal of these "brakes" in T cells results in increased activation of the immune system and controlling and eradicating tumor. The development of immune checkpoint inhibitors (ICI...

journal_title：Methods in enzymology

authors： Nomura N,Nomura Y,Sato Y,Iwata S

更新日期：2019-01-01 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：:In recent years the facile, yet powerful, genetics of the baker's yeast Saccharomyces cerevisiae has been appropriated for the study of amyloid toxicity. Several models of amyloid toxicity using this simple eukaryotic organism have been developed that faithfully recapitulate many disease-relevant phenotypes. Furthermo...

journal_title：Methods in enzymology

authors： Giorgini F,Muchowski PJ

更新日期：2006-01-01 00:00:00

abstract：:Once the appropriate site has been selected for the attachment of GFP to the sarcomeric protein, it is quite remarkable that the large size of the GFP molecule does not appear to interfere with the localization of the fluorescent sarcomeric proteins into the sarcomeric regions of the myofibrils. A similar approach usi...

journal_title：Methods in enzymology

authors： Dabiri GA,Ayoob JC,Turnacioglu KK,Sanger JM,Sanger JW

更新日期：1999-01-01 00:00:00

abstract：:Bispecific antibodies may be used to improve clinical efficacy by targeting two disease mechanisms for the treatment of complex human diseases in a single agent. Bispecific antibodies also hold promise for certain therapeutic applications difficult to achieve by single-targeting monospecific antibodies, such as immune...

journal_title：Methods in enzymology

authors： Gu J,Ghayur T

更新日期：2012-01-01 00:00:00