On-pot and cell-free biocatalysis using coimmobilized enzymes on advanced materials.

abstract：:The remarkable success of cancer immunotherapies, especially the checkpoint blocking antibodies, in a subset of patients has reinvigorated the study of tumor-immune crosstalk and its role in heterogeneity of response. High-throughput sequencing and imaging technologies can help recapitulate various aspects of the tumo...

journal_title：Methods in enzymology

authors： Bhinder B,Elemento O

更新日期：2020-01-01 00:00:00

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：:The analysis of a reaction pathway by stopped-flow and chemical quench-flow methods provides the only means of directly measuring the rates of reaction at the active site of an enzyme. The interpretation of data from each technique is similar and involves solution of the differential equations describing the approach ...

journal_title：Methods in enzymology

authors： Johnson KA

更新日期：1986-01-01 00:00:00

abstract：:Neuroblastoma (NB) is the most common extracranial solid tumor in childhood and the most frequently diagnosed neoplasm during infancy. Despite of aggressive treatment strategies, the 5-year survival rate for metastatic disease is still less than 60% and, consequently, novel therapeutic approaches are needed. For incre...

journal_title：Methods in enzymology

authors： Di Paolo D,Loi M,Pastorino F,Brignole C,Marimpietri D,Becherini P,Caffa I,Zorzoli A,Longhi R,Gagliani C,Tacchetti C,Corti A,Allen TM,Ponzoni M,Pagnan G

更新日期：2009-01-01 00:00:00

abstract：:The ability to examine gene regulation in living cells has been greatly enabled by the development of chromatin immunoprecipitation (ChIP) methodology. ChIP captures a snapshot of protein-DNA interactions in vivo and has been used to study interactions in bacteria, yeast, and mammalian cell culture. ChIP conditions va...

journal_title：Methods in enzymology

authors： Davis SE,Mooney RA,Kanin EI,Grass J,Landick R,Ansari AZ

更新日期：2011-01-01 00:00:00

abstract：:Osmoregulation is the active control of the cellular water balance and encompasses homeostatic mechanisms crucial for life. The osmoregulatory system in the yeast Saccharomyces cerevisiae is particularly well understood. Key to yeast osmoregulation is the production and accumulation of the compatible solute glycerol, ...

journal_title：Methods in enzymology

authors： Hohmann S,Krantz M,Nordlander B

更新日期：2007-01-01 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：:In budding yeast and higher eukaryotic genomes, there are, respectively, 50 and up to 400 or more distinct genes that encode for ubiquitin-ligases, and approximately 15-90 genes that encode for ubiquitin isopeptidases (TM and RJD, Semple et al., 2003). This puts ubiquitylation on par with phosphorylation as the most c...

journal_title：Methods in enzymology

authors： Mayor T,Deshaies RJ

更新日期：2005-01-01 00:00:00

abstract：:This protocol describes a method for determining the concentration of a nucleic acid sample. ...

journal_title：Methods in enzymology

authors： Kolitz S,Lorsch J

更新日期：2013-01-01 00:00:00

abstract：:The enzymes involved in mitochondrial oxidative phosphorylation (OXPHOS) are coassembled into higher ordered supercomplexes within the mitochondrial inner membrane. The cytochrome bc(1)-cytochrome c oxidase (COX) supercomplex is formed by the coassociation of the two electron transport chain complexes, the cytochrome ...

journal_title：Methods in enzymology

authors： Stuart RA

更新日期：2009-01-01 00:00:00

abstract：:Oxidized phospholipids play an important role in execution of the mitochondrial stage of apoptosis and clearance of apoptotic cells by macrophages. Therefore, the identification and quantification of oxidized phospholipids generated during apoptosis are very important. These can be achieved successfully by a newly dev...

journal_title：Methods in enzymology

authors： Tyurin VA,Tyurina YY,Kochanek PM,Hamilton R,DeKosky ST,Greenberger JS,Bayir H,Kagan VE

更新日期：2008-01-01 00:00:00

abstract：:Human embryonic stem cells hold great promise in furthering our treatment of disease and increasing our understanding of early development. This chapter describes protocols for the derivation and maintenance of human embryonic stem cells. In addition, it summarizes briefly several alternative methods for the culture o...

journal_title：Methods in enzymology

authors： Akutsu H,Cowan CA,Melton D

更新日期：2006-01-01 00:00:00

abstract：:Fertilization by intracytoplasmic sperm injection (ICSI) is a powerful technique that can be used to understand better the biology of fertilization, in addition to a form of assisted reproduction both in humans and in endangered species. Mouse is often the model organism of choice to study mammalian fertilization. The...

journal_title：Methods in enzymology

authors： Stein P,Schultz RM

更新日期：2010-01-01 00:00:00

abstract：:Acidic organelles and vesicles, such as endosomes, lysosomes, autophagosomes, trans-Golgi network, and synaptic vesicles, are known to play important roles in a broad range of cellular events. To facilitate studying these multifunctional systems, we describe here an acid-brightening fluorescent protein (abFP), which f...

journal_title：Methods in enzymology

authors： Wang N,Wang L

更新日期：2020-01-01 00:00:00

abstract：:Protein oxidation is implicated in atherogenesis and other inflammatory conditions. Measuring levels of chlorinated and nitrated proteins in biological matrices serves as a quantitative index of oxidative stress in vivo. One potential mechanism for oxidative stress involves myeloperoxidase, a heme protein expressed by...

journal_title：Methods in enzymology

authors： Shao B,Heinecke JW

更新日期：2008-01-01 00:00:00

abstract：:Tumor cells acquire distinct genetic characteristics as a means to survive and proliferate indefinitely. Changes in the genetic code can also translate in changes at the protein level, therefore creating a distinguishable signature unique for tumor cells, and absent in normal tissues. The presence of discernable moiet...

journal_title：Methods in enzymology

authors： Battaglia S

更新日期：2020-01-01 00:00:00

abstract：:Amyloid aggregates are associated with a number of mammalian neurodegenerative diseases. Infectious aggregates of the mammalian prion protein PrP(sc) are hallmarks of transmissible spongiform encephalopathies in humans and cattle (Griffith, 1967; Legname et al., 2004; Prusiner, 1982; Silveira et al., 2004). Likewise, ...

journal_title：Methods in enzymology

authors： Bagriantsev SN,Kushnirov VV,Liebman SW

更新日期：2006-01-01 00:00:00

abstract：:Regulator of G-protein signaling (RGS) domains bind directly to GTP-bound Galpha subunits and accelerate their intrinsic GTPase activity by up to several thousandfold. The selectivity of RGS proteins for individual Galpha subunits has been illustrated. Thus, the expression of RGS proteins can be used to inhibit signal...

journal_title：Methods in enzymology

authors： Hains MD,Siderovski DP,Harden TK

更新日期：2004-01-01 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：:Glass microfluidic devices have been fabricated to monitor the secretion of glycerol or fatty acids from cultured murine 3T3-L1 adipocytes. In the current studies, adipocytes are perfused in a reversibly sealed cell chamber, and secreted products are analyzed by enzyme assay on either a single- or dual-chip device. Th...

journal_title：Methods in enzymology

authors： Dugan CE,Kennedy RT

更新日期：2014-01-01 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：:Guanine nucleotide exchange factors (GEFs) are essential regulators of the spatiotemporal conditions of small GTP-binding protein (SMG) activation. Their cellular activities combine the biochemical stimulation of GDP/GTP exchange, which leads to the active conformation of the SMG, to the detection of upstream signals ...

journal_title：Methods in enzymology

authors： Zeeh JC,Antonny B,Cherfils J,Zeghouf M

更新日期：2008-01-01 00:00:00

abstract：:We have used a shortened construct form of the CYP26A1 gene promoter, in a promoter-less vector with either luciferase (known as E4) or a red fluorescent protein, RFP (known as E4.2) as the reporter gene and examined their responses to retinoids in transfected HepG2 and HEK293T cells. The promoter responded linearly t...

journal_title：Methods in enzymology

authors： Zolfaghari R,Mattie FJ,Wei CH,Chisholm DR,Whiting A,Ross AC

更新日期：2020-01-01 00:00:00

abstract：:Heterodimeric complexes between individual members of the R7 subfamily of regulators of G-protein signaling proteins and the Gbeta5 isoform of the heterotrimeric G-protein beta subunit family are strongly expressed in the cell nucleus in neurons and brain, as well as in the cytoplasm and plasma membrane. Native and re...

journal_title：Methods in enzymology

authors： Simonds WF,Woodard GE,Zhang JH

更新日期：2004-01-01 00:00:00

abstract：:Graphene oxide (GO) and reduced graphene oxide (rGO) were demonstrated in the past decade as biocompatible carbon-based materials that could be efficiently used in bioelectrochemical systems (BESs). Specifically, for redox enzyme encapsulation in order to improve electron communication between enzymes and electrodes. ...

journal_title：Methods in enzymology

authors： Schlesinger O,Alfonta L

更新日期：2018-01-01 00:00:00

abstract：:Aging that involves contact with dying yeast cells is one of the differential processes between sparkling and still wine production. The release of the products of autolysis during this aging step is fundamental for the quality of sparkling wines made by the traditional method. These cells undergo an autolysis process...

journal_title：Methods in enzymology

authors： Cebollero E,Rejas MT,González R

更新日期：2008-01-01 00:00:00

abstract：:5-FOA is an extremely useful reagent for the selection of Ura- cells amid a population of Ura+ cells. The selection is effective in transformation and recombination studies where loss of URA3+ is desired. A new plasmid shuffling procedure based on the 5-FOAR selection permits the recovery of conditional lethal mutatio...

journal_title：Methods in enzymology

authors： Boeke JD,Trueheart J,Natsoulis G,Fink GR

更新日期：1987-01-01 00:00:00

abstract：:Acetyl CoA:arylamine N-acetyltransferase (NAT; E.C. 2.3.1.5) enzymes play a key role in the metabolic activation of aromatic amine and nitroaromatic mutagens to electrophilic reactive intermediates. We have developed a system in which the activation of mutagens by recombinant human NAT2, expressed in Escherichia coli,...

journal_title：Methods in enzymology

authors： Savulescu MR,Mushtaq A,Josephy PD

更新日期：2005-01-01 00:00:00

abstract：:Glycosaminoglycans (GAGs) are long unbranched polysaccharides, most of which are linked to a core protein to form proteoglycans. Depending on the nature of their backbone, one can discern galactosaminoglycans (chondroitin sulfate [CS] and dermatan sulfate [DS]) and glucosaminoglycans (heparan sulfate [HS], heparin, hy...

journal_title：Methods in enzymology

authors： Smits NC,Lensen JF,Wijnhoven TJ,Ten Dam GB,Jenniskens GJ,van Kuppevelt TH

更新日期：2006-01-01 00:00:00

abstract：:The histamine H(1) receptor (H(1)R) is a key player in acute inflammatory responses. Antihistamines are widely used to relief the symptoms of allergic rhinitis by antagonizing histamine binding to the H(1)R, without possessing intrinsic activity in classical assays such as guinea pig ileum contraction assays or intrac...

journal_title：Methods in enzymology

authors： Nijmeijer S,Leurs R,Vischer HF

更新日期：2010-01-01 00:00:00