Reconstitution of the cell cycle-regulated Golgi disassembly and reassembly in a cell-free system.

abstract：:The new generation of synchrotrons and microfocused beamlines has enabled great progress in X-ray protein crystallography, resulting in new 3D atomic structures for proteins of high interest to the pharmaceutical industry and life sciences. It is, however, often still challenging to produce protein crystals of suffici...

journal_title：Nature protocols

authors： Pechkova E,Nicolini C

更新日期：2017-12-01 00:00:00

abstract：:We describe a simple and versatile scheme to prepare a series of poly(ethylene glycol)-based bidentate ligands that permit strong interactions with colloidal semiconductor nanocrystals (quantum dots, QDs) and gold nanoparticles (AuNPs) alike and promote their dispersion in aqueous solutions. These ligands are synthesi...

journal_title：Nature protocols

authors： Mei BC,Susumu K,Medintz IL,Mattoussi H

更新日期：2009-01-01 00:00:00

abstract：:Several cell death assays have been developed based on a single biochemical parameter such as caspase activation or plasma membrane permeabilization. Our fluorescent apoptosis/necrosis (FAN) assay directly measures cell death and distinguishes between caspase-dependent apoptosis and caspase-independent necrosis of cel...

journal_title：Nature protocols

authors： Grootjans S,Hassannia B,Delrue I,Goossens V,Wiernicki B,Dondelinger Y,Bertrand MJ,Krysko DV,Vuylsteke M,Vandenabeele P,Vanden Berghe T

更新日期：2016-08-01 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：:In most organisms, one crossover (CO) event inhibits the chances of another nearby event. The term used to describe this phenomenon is 'CO interference'. Here, we describe a protocol for quickly generating large data sets that are amenable to CO interference analysis in the flowering plant, Arabidopsis thaliana. We em...

journal_title：Nature protocols

authors： Berchowitz LE,Copenhaver GP

更新日期：2008-01-01 00:00:00

abstract：:The analysis of highly hydrophobic proteins is still an analytical challenge. Using a recombinant gamma-aminobutyric acid A (GABAA)-receptor subunit as a model protein, we developed a gel-based proteomic approach for high MS/MS-peptide sequence coverage identification. Protein samples were separated by multi-dimension...

journal_title：Nature protocols

authors： Kang SU,Fuchs K,Sieghart W,Pollak A,Csaszar E,Lubec G

更新日期：2009-01-01 00:00:00

abstract：:This protocol outlines the carboxyfluorescein diacetate succinimidyl ester (CFSE) method for following the proliferation of human lymphocytes in vitro and mouse lymphocytes both in vitro and in vivo. The method relies on the ability of CFSE to covalently label long-lived intracellular molecules with the highly fluores...

journal_title：Nature protocols

authors： Quah BJ,Warren HS,Parish CR

更新日期：2007-01-01 00:00:00

abstract：:We describe the implementation and use of an adaptive imaging framework for optimizing spatial resolution and signal strength in a light-sheet microscope. The framework, termed AutoPilot, comprises hardware and software modules for automatically measuring and compensating for mismatches between light-sheet and detecti...

journal_title：Nature protocols

authors： Royer LA,Lemon WC,Chhetri RK,Keller PJ

更新日期：2018-11-01 00:00:00

abstract：:This protocol provides a detailed procedure for the chemical synthesis of proteins through native chemical ligation of peptide hydrazides. The two crucial stages of this protocol are (i) the solid-phase synthesis of peptide hydrazides via Fmoc chemistry and (ii) the native chemical ligation of peptide hydrazides throu...

journal_title：Nature protocols

authors： Zheng JS,Tang S,Qi YK,Wang ZP,Liu L

更新日期：2013-12-01 00:00:00

abstract：:The immune system operates at the scale of the whole organism in mammals. We currently lack experimental approaches to systematically track and study organism-wide molecular processes in mice. Here we describe an integrated toolkit for measuring gene expression in whole tissues, 3-prime mRNA extension sequencing, that...

journal_title：Nature protocols

authors： Pandey S,Takahama M,Gruenbaum A,Zewde M,Cheronis K,Chevrier N

更新日期：2020-04-01 00:00:00

abstract：:Reversible thiol oxidation of cysteine residues occurs in many intracellular catalytic and signaling processes. Here we describe an optimized protocol, which can be completed in ∼5 d, to unambiguously identify specific cysteine residues that are transiently and reversibly oxidized by comparing two complex biological s...

journal_title：Nature protocols

authors： García-Santamarina S,Boronat S,Domènech A,Ayté J,Molina H,Hidalgo E

更新日期：2014-05-01 00:00:00

abstract：:The rodent malaria parasite Plasmodium chabaudi chabaudi shares many features with human malaria species, including P. falciparum, and is the in vivo model of choice for many aspects of malaria research in the mammalian host, from sequestration of parasitized erythrocytes, to antigenic variation and host immunity and ...

journal_title：Nature protocols

authors： Spence PJ,Cunningham D,Jarra W,Lawton J,Langhorne J,Thompson J

更新日期：2011-04-01 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：: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：:Blood circulating endothelial cells (CECs) and circulating hematopoietic progenitor cells (CPCs) represent two cell populations that are thought to play important roles in tissue vascularization. CECs and CPCs are currently studied as surrogate markers in patients for more than a dozen pathologies, including heart dis...

journal_title：Nature protocols

authors： Duda DG,Cohen KS,Scadden DT,Jain RK

更新日期：2007-01-01 00:00:00

abstract：:RNA sequencing (RNA-seq) has been rapidly adopted for the profiling of transcriptomes in many areas of biology, including studies into gene regulation, development and disease. Of particular interest is the discovery of differentially expressed genes across different conditions (e.g., tissues, perturbations) while opt...

journal_title：Nature protocols

authors： Anders S,McCarthy DJ,Chen Y,Okoniewski M,Smyth GK,Huber W,Robinson MD

更新日期：2013-09-01 00:00:00

abstract：:A big challenge in proteomics is the identification of cell-type-specific proteomes in vivo. This protocol describes how to label, purify and identify cell-type-specific proteomes in living mice. To make this possible, we created a Cre-recombinase-inducible mouse line expressing a mutant methionyl-tRNA synthetase (L27...

journal_title：Nature protocols

authors： Alvarez-Castelao B,Schanzenbächer CT,Langer JD,Schuman EM

更新日期：2019-02-01 00:00:00

abstract：:Electrochemically active bacteria (EAB) have the ability to transfer electrons to electron acceptors located outside the cell, and they are widely present in diverse environments. In spite of their important roles in geochemical cycles, environmental remediation and electricity generation, so far, only a limited numbe...

journal_title：Nature protocols

authors： Yuan SJ,Li WW,Cheng YY,He H,Chen JJ,Tong ZH,Lin ZQ,Zhang F,Sheng GP,Yu HQ

更新日期：2014-01-01 00:00:00

abstract：:Accurate analysis of DNA sequence variation in not only humans and animals but also other organisms has played a significant role in expanding our knowledge about genetic variety and diversity in a number of different biological areas. The search for an understanding of the causes of genetic variants and mutations has...

journal_title：Nature protocols

authors： Ota M,Fukushima H,Kulski JK,Inoko H

更新日期：2007-01-01 00:00:00

abstract：:Periodontal disease (PD) is a common dental disease associated with the interaction between dysbiotic oral microbiota and host immunity. It is a prevalent disease, resulting in loss of gingival tissue, periodontal ligament, cementum and alveolar bone. PD is a major form of tooth loss in the adult population. Experimen...

journal_title：Nature protocols

authors： Marchesan J,Girnary MS,Jing L,Miao MZ,Zhang S,Sun L,Morelli T,Schoenfisch MH,Inohara N,Offenbacher S,Jiao Y

更新日期：2018-10-01 00:00:00

abstract：:In this protocol, we describe cryoimmunolabeling methods for the subcellular localization of proteins and certain lipids. The methods start with chemical fixation of cells and tissue in formaldehyde (FA) and/or glutaraldehyde (GA), sometimes supplemented with acrolein. Cell and tissue blocks are then immersed in 2.3 M...

journal_title：Nature protocols

authors： Slot JW,Geuze HJ

更新日期：2007-01-01 00:00:00

abstract：:Although differences in protein staining intensity can often be visualized by difference gel electrophoresis, abundant proteins can obscure less abundant proteins, and quantification of post-translational modifications is difficult. We present a protocol for quantifying changes in the abundance of a specific protein o...

journal_title：Nature protocols

authors： Asara JM,Zhang X,Zheng B,Maroney LA,Christofk HR,Wu N,Cantley LC

更新日期：2006-01-01 00:00:00

abstract：:This protocol represents a novel enzyme-luminescence method to detect dopamine sensitively and rapidly with high temporal resolution. In principle, dopamine is first oxidized with tyramine oxidase to produce H(2)O(2), and then the produced H(2)O(2) reacts with luminol to generate chemiluminescence in the presence of h...

journal_title：Nature protocols

authors： Shinohara H,Wang F,Hossain SM

更新日期：2008-01-01 00:00:00

abstract：:The glycocalyx comprises glycosylated proteins and lipids and fcorms the outermost layer of cells. It is involved in fundamental inter- and intracellular processes, including non-self-cell and self-cell recognition, cell signaling, cellular structure maintenance, and immune protection. Characterization of the glycocal...

journal_title：Nature protocols

authors： Li Q,Xie Y,Wong M,Barboza M,Lebrilla CB

更新日期：2020-08-01 00:00:00

abstract：:Analysis of the sequence and nature of protein N termini has many applications. Defining the termini of proteins for proteome annotation in the Human Proteome Project is of increasing importance. Terminomics analysis of protease cleavage sites in degradomics for substrate discovery is a key new application. Here we de...

journal_title：Nature protocols

authors： Kleifeld O,Doucet A,Prudova A,auf dem Keller U,Gioia M,Kizhakkedathu JN,Overall CM

更新日期：2011-09-22 00:00:00

abstract：:Protein ubiquitination is a versatile protein modification that regulates virtually all cellular processes. This versatility originates from polyubiquitin chains, which can be linked in eight distinct ways. The combinatorial complexity of eight linkage types in homotypic (one chain type per polymer) and heterotypic (m...

journal_title：Nature protocols

authors： Hospenthal MK,Mevissen TET,Komander D

更新日期：2015-02-01 00:00:00

abstract：:Gene tagging with fluorescent proteins is essential for investigations of the dynamic properties of cellular proteins. CRISPR-Cas9 technology is a powerful tool for inserting fluorescent markers into all alleles of the gene of interest (GOI) and allows functionality and physiological expression of the fusion protein. ...

journal_title：Nature protocols

authors： Koch B,Nijmeijer B,Kueblbeck M,Cai Y,Walther N,Ellenberg J

更新日期：2018-06-01 00:00:00

abstract：:Interactions of proteins with DNA mediate many critical nuclear functions. Chromatin immunoprecipitation (ChIP) is a robust technique for studying protein-DNA interactions. Current ChIP assays, however, either require large cell numbers, which prevent their application to rare cell samples or small-tissue biopsies, or...

journal_title：Nature protocols

authors： Dahl JA,Collas P

更新日期：2008-01-01 00:00:00

abstract：:The goal of many current studies of neovascularization is to define the phenotype of vascular cell populations of different origins and to determine how such cells promote assembly of vascular channel. Here, we describe a protocol to immunophenotype vascular cells by high-resolution imaging and by fluorescence-activat...

journal_title：Nature protocols

authors： Jones RC,Capen DE,Cohen KS,Munn LL,Jain RK,Duda DG

更新日期：2008-01-01 00:00:00

abstract：:The CRISPR-Cas9 genome-editing tool and the availability of whole-genome sequences from plant species have revolutionized our ability to introduce targeted mutations into important crop plants, both to explore genetic changes and to introduce new functionalities. Here, we describe protocols adapting the CRISPR-Cas9 sy...

journal_title：Nature protocols

authors： Osakabe Y,Liang Z,Ren C,Nishitani C,Osakabe K,Wada M,Komori S,Malnoy M,Velasco R,Poli M,Jung MH,Koo OJ,Viola R,Nagamangala Kanchiswamy C

更新日期：2018-12-01 00:00:00