Analysis of mutational spectra by denaturing capillary electrophoresis.

abstract：:Gram-negative and Gram-positive bacteria release a variety of membrane vesicles through different formation routes. Knowledge of the structure, molecular cargo and function of bacterial extracellular vesicles (BEVs) is primarily obtained from bacteria cultured in laboratory conditions. BEVs in human body fluids have b...

journal_title：Nature protocols

authors： Tulkens J,De Wever O,Hendrix A

更新日期：2020-01-01 00:00:00

abstract：:The accurate knowledge of receptor kinetics is crucial to our understanding of cell signal transduction in general and neural function in particular. The classical technique of probing membrane receptors on a millisecond scale involves placing a recording micropipette with a membrane patch in front of a double-barrel ...

journal_title：Nature protocols

authors： Sylantyev S,Rusakov DA

更新日期：2013-01-01 00:00:00

abstract：:In this report, we describe a two-step protocol for labeling of an affinity-purified antibody to biotin with horseradish peroxidase (HRP) using cyanuric chloride (CC) as a bridge. The enzyme was first modified with CC, and following chromatography on a PD-10 column, the activated HRP was incubated with the antibody to...

journal_title：Nature protocols

authors： Abuknesha RA,Jeganathan F,Wu J,Baalawy Z

更新日期：2009-01-01 00:00:00

abstract：:Apoptosis is a critical factor in AIDS and other viral illnesses, cerebral and myocardial ischemia, autoimmune and neurodegenerative states, organ and bone marrow transplant rejection, and tumor response to chemotherapy and radiation. Improved methods to identify sites of apoptosis are increasing our understanding of ...

journal_title：Nature protocols

authors： Blankenberg FG,Vanderheyden JL,Strauss HW,Tait JF

更新日期：2006-01-01 00:00:00

abstract：:Optogenetic tools provide users the ability to photocontrol the activity of cells. Commonly, activation is achieved by expression of proteins from photosynthetic organisms, for example, microbial opsins (e.g., ChR2). Alternatively, a sister approach, synthetic optogenetics, enables photocontrol over proteins of mammal...

journal_title：Nature protocols

authors： Carmi I,De Battista M,Maddalena L,Carroll EC,Kienzler MA,Berlin S

更新日期：2019-03-01 00:00:00

abstract：:Live single-cell mass spectrometry (live MS) provides a mass spectrum that shows thousands of metabolite peaks from a single live plant cell within minutes. By using an optical microscope, a cell is chosen for analysis and a metal-coated nanospray microcapillary tip is used to remove the cell's contents. After adding ...

journal_title：Nature protocols

authors： Fujii T,Matsuda S,Tejedor ML,Esaki T,Sakane I,Mizuno H,Tsuyama N,Masujima T

更新日期：2015-09-01 00:00:00

abstract：:N-Succinimidyl 3-(di-tert-butyl[(18)F]fluorosilyl)benzoate ([(18)F]SiFB) is a highly reactive prosthetic group for radiolabeling of proteins for use in positron emission tomography (PET). It is similar to N-succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB), the 'gold-standard' prosthetic group for protein (18)F-labeli...

journal_title：Nature protocols

authors： Kostikov AP,Chin J,Orchowski K,Schirrmacher E,Niedermoser S,Jurkschat K,Iovkova-Berends L,Wängler C,Wängler B,Schirrmacher R

更新日期：2012-11-01 00:00:00

abstract：:The tracking of cell fate, shape and migration is an essential component in the study of the development of multicellular organisms. Here we report a protocol that uses the protein Kaede, which is fluorescent green after synthesis but can be photoconverted red by violet or UV light. We have used Kaede along with confo...

journal_title：Nature protocols

authors： Hatta K,Tsujii H,Omura T

更新日期：2006-01-01 00:00:00

abstract：:Establishing a small animal model that accurately recapitulates hepatotropic pathogens, including hepatitis C virus (HCV) infection and immunopathogenesis, is essential for the study of hepatitis virus-induced liver disease and for therapeutics development. This protocol describes our recently developed humanized mous...

journal_title：Nature protocols

authors： Bility MT,Zhang L,Washburn ML,Curtis TA,Kovalev GI,Su L

更新日期：2012-09-01 00:00:00

abstract：:The use of anisotropic NMR data, such as residual dipolar couplings (RDCs) and residual chemical shift anisotropies (RCSAs), has emerged as a powerful technique for structural characterization of organic small molecules. RDCs typically report the relative orientations of different 1H-13C bonds; RCSAs report the relati...

journal_title：Nature protocols

authors： Liu Y,Navarro-Vázquez A,Gil RR,Griesinger C,Martin GE,Williamson RT

更新日期：2019-01-01 00:00:00

abstract：:Genetically encoded fluorescent proteins (FPs) are highly utilized in cell biology research to study proteins of interest or signal processes using biosensors. To perform well in specific applications, these FPs require a multitude of tailored properties. It is for this reason that they need to be optimized by using m...

journal_title：Nature protocols

authors： Bindels DS,Postma M,Haarbosch L,van Weeren L,Gadella TWJ Jr

更新日期：2020-02-01 00:00:00

abstract：:An optimized procedure for the efficient preparation of 2-heptyl-3-hydroxy-4(1H)-quinolone (Pseudomonas quinolone signal or PQS) and a diverse range of structurally related 2-alkyl-4-quinolones with biological activity is presented. The two-step synthesis begins with the formation of α-chloro ketones by the coupling o...

journal_title：Nature protocols

authors： Hodgkinson JT,Galloway WR,Welch M,Spring DR

更新日期：2012-05-24 00:00:00

abstract：:Social recognition (SR) enables rodents to distinguish between familiar and novel conspecifics, largely through individual odor cues. SR tasks utilize the tendency for a male to sniff and interact with a novel individual more than a familiar individual. Many paradigms have been used to study the roles of the neuropept...

journal_title：Nature protocols

authors： Macbeth AH,Edds JS,Young WS 3rd

更新日期：2009-01-01 00:00:00

abstract：:The dynamics of transcription can be studied genome wide by high-throughput sequencing of nascent and newly synthesized RNA. 4-thiouridine (4SU) labeling in vivo enables the specific capture of such new transcripts, with 4SU residues being tagged by biotin linkers and captured using streptavidin beads before library p...

journal_title：Nature protocols

authors： Gregersen LH,Mitter R,Svejstrup JQ

更新日期：2020-02-01 00:00:00

abstract：:Brachypodium distachyon is emerging as a new model system for bridging research into temperate cereal crops, such as wheat and barley, and for promoting research in novel biomass grasses. Here, we provide an adapter ligation PCR protocol that allows the large-scale characterization of T-DNA insertions into the genome ...

journal_title：Nature protocols

authors： Thole V,Alves SC,Worland B,Bevan MW,Vain P

更新日期：2009-01-01 00:00:00

abstract：:The ability to rapidly generate large panels of antigen-specific human antibodies in a rodent would enable the efficient discovery of novel therapeutically useful antibodies. We have developed a system wherein human antigen-specific antibody-secreting plasmablasts can be enriched in vivo, in a severe combined immunode...

journal_title：Nature protocols

authors： Lin Z,Chiang NY,Chai N,Seshasayee D,Lee WP,Balazs M,Nakamura G,Swem LR

更新日期：2014-07-01 00:00:00

abstract：:Mitophagy is a cellular process that selectively removes damaged, old or dysfunctional mitochondria. Defective mitophagy is thought to contribute to normal aging and to various neurodegenerative and cardiovascular diseases. Previous methods used to detect mitophagy in vivo were cumbersome, insensitive and difficult to...

journal_title：Nature protocols

authors： Sun N,Malide D,Liu J,Rovira II,Combs CA,Finkel T

更新日期：2017-08-01 00:00:00

abstract：:The cytokinesis-block micronucleus cytome assay is a comprehensive system for measuring DNA damage, cytostasis and cytotoxicity. DNA damage events are scored specifically in once-divided binucleated (BN) cells and include (a) micronuclei (MNi), a biomarker of chromosome breakage and/or whole chromosome loss, (b) nucle...

journal_title：Nature protocols

authors： Fenech M

更新日期：2007-01-01 00:00:00

abstract：:Carbohydrate microarrays have received considerable attention as an advanced technology for the rapid analysis of carbohydrate-protein interactions. This protocol provides detailed procedures for the preparation of carbohydrate microarrays by immobilizing hydrazide-conjugated carbohydrates on epoxide-derivatized glass...

journal_title：Nature protocols

authors： Park S,Lee MR,Shin I

更新日期：2007-01-01 00:00:00

abstract：:Here, we provide comprehensive, highly efficient protocols for Agrobacterium tumefaciens-mediated transformation of a wide range of rice genotypes. Methods that use either immature embryos (japonica and indica rice) or calli (japonica cultivars and the indica cultivar, Kasalath) as a starting material for inoculation ...

journal_title：Nature protocols

authors： Hiei Y,Komari T

更新日期：2008-01-01 00:00:00

abstract：:A mechanistic understanding of HIV-1 latency depends on a model system that recapitulates the in vivo condition of latently infected, resting CD4(+) T lymphocytes. Latency seems to be established after activated CD4(+) T cells, the principal targets of HIV-1 infection, become productively infected and survive long eno...

journal_title：Nature protocols

authors： Kim M,Hosmane NN,Bullen CK,Capoferri A,Yang HC,Siliciano JD,Siliciano RF

更新日期：2014-12-01 00:00:00

abstract：:The pollen tube is an excellent single-cell model system for studying cellular processes in plant cell biology. This protocol describes a detailed step-by-step procedure with optimized conditions for introducing various fluorescent reporter proteins into lily, tobacco and Arabidopsis pollen grains by means of biolisti...

journal_title：Nature protocols

authors： Wang H,Jiang L

更新日期：2011-04-01 00:00:00

abstract：:This protocol describes a method for direct labeling and detection of small RNAs present in total RNA by splinted ligation. The assay uses a small RNA-specific bridge oligonucleotide to form base pairs with the small RNA and a 5'-end-radiolabeled ligation oligonucleotide. The captured small RNA is directly labeled by ...

journal_title：Nature protocols

authors： Maroney PA,Chamnongpol S,Souret F,Nilsen TW

更新日期：2008-01-01 00:00:00

abstract：:Here we present a protocol for the extraction of RNA from Dictyostelium discoideum. Dictyostelium is a social amoeba that undergoes a basic developmental program, and therefore analysis of RNA levels over a time course is a commonly used technique. This procedure is similar to other guanidine thiocyanate-based methods...

journal_title：Nature protocols

authors： Pilcher KE,Gaudet P,Fey P,Kowal AS,Chisholm RL

更新日期：2007-01-01 00:00:00

abstract：:Aptamers are single-stranded DNA or RNA molecules that can bind target molecules with high affinity and specificity. The conformation of an aptamer usually changes upon binding to its target analyte, and this property has been used in a wide variety of sensing applications, including detection based on fluorescence in...

journal_title：Nature protocols

authors： Liu J,Lu Y

更新日期：2006-01-01 00:00:00

abstract：:Single-cell electroporation allows transfection of plasmid DNA or macromolecules into individual living cells using modified patch electrodes and common electrophysiological equipment. This protocol is optimized for rapid in vivo electroporation of Xenopus laevis tadpole brains with DNA, dextrans, morpholinos and comb...

journal_title：Nature protocols

authors： Bestman JE,Ewald RC,Chiu SL,Cline HT

更新日期：2006-01-01 00:00:00

abstract：:Targeted nucleases, including zinc-finger nucleases (ZFNs), transcription activator-like (TAL) effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9), have provided researchers with the ability to manipulate nearly any genomic sequence in hu...

journal_title：Nature protocols

authors： Liu J,Gaj T,Yang Y,Wang N,Shui S,Kim S,Kanchiswamy CN,Kim JS,Barbas CF 3rd

更新日期：2015-11-01 00:00:00

abstract：:Cell migration through 3D extracellular matrices (ECMs) is crucial to the normal development of tissues and organs and in disease processes, yet adequate analytical tools to characterize 3D migration are lacking. The motility of eukaryotic cells on 2D substrates in the absence of gradients has long been described usin...

journal_title：Nature protocols

authors： Wu PH,Giri A,Wirtz D

更新日期：2015-03-01 00:00:00

abstract：:We describe the use of a microfabricated cell culture substrate, consisting of a uniform array of closely spaced, vertical, elastomeric microposts, to study the effects of substrate rigidity on cell function. Elastomeric micropost substrates are micromolded from silicon masters comprised of microposts of different hei...

journal_title：Nature protocols

authors： Yang MT,Fu J,Wang YK,Desai RA,Chen CS

更新日期：2011-02-01 00:00:00

abstract：:Multipotent neural crest stem cells (NCSCs) have the potential to generate a wide range of cell types including melanocytes; peripheral neurons; and smooth muscle, bone, cartilage and fat cells. This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent c...

journal_title：Nature protocols

authors： Menendez L,Kulik MJ,Page AT,Park SS,Lauderdale JD,Cunningham ML,Dalton S

更新日期：2013-01-01 00:00:00