Genetic transformation of tobacco NT1 cells with Agrobacterium tumefaciens.

abstract：:Forebrain γ-aminobutyric acid (GABA) interneurons have crucial roles in high-order brain function via modulating network activities and plasticity, and they are implicated in many psychiatric disorders. Availability of enriched functional human forebrain GABA interneurons, especially those from people affected by GABA...

journal_title：Nature protocols

authors： Liu Y,Liu H,Sauvey C,Yao L,Zarnowska ED,Zhang SC

更新日期：2013-09-01 00:00:00

abstract：:Using siRNAs to genetically manipulate immune cells is important to both basic immunological studies and therapeutic applications. However, siRNA delivery is challenging because primary immune cells are often sensitive to the delivery materials and generate immune responses. We have recently developed an amphiphilic d...

journal_title：Nature protocols

authors： Chen J,Ellert-Miklaszewska A,Garofalo S,Dey AK,Tang J,Jiang Y,Clément F,Marche PN,Liu X,Kaminska B,Santoni A,Limatola C,Rossi JJ,Zhou J,Peng L

更新日期：2021-01-01 00:00:00

abstract：:The organization of eukaryotic cells into distinct subcompartments is vital for all functional processes, and aberrant protein localization is a hallmark of many diseases. Microscopy methods, although powerful, are usually low-throughput and dependent on the availability of fluorescent fusion proteins or highly specif...

journal_title：Nature protocols

authors： Mulvey CM,Breckels LM,Geladaki A,Britovšek NK,Nightingale DJH,Christoforou A,Elzek M,Deery MJ,Gatto L,Lilley KS

更新日期：2017-06-01 00:00:00

abstract：:This protocol describes a simple but robust microfluidic assay combining three-dimensional (3D) and two-dimensional (2D) cell culture. The microfluidic platform comprises hydrogel-incorporating chambers between surface-accessible microchannels. By using this platform, well-defined biochemical and biophysical stimuli c...

journal_title：Nature protocols

authors： Shin Y,Han S,Jeon JS,Yamamoto K,Zervantonakis IK,Sudo R,Kamm RD,Chung S

更新日期：2012-06-07 00:00:00

abstract：:We describe a step-by-step protocol for measuring the stable products of the nitric oxide (NO) pathway: nitrite, nitrite plus nitrate and nitrate. This described protocol is easy to apply and is about 50 times more sensitive than the commonly used Griess reaction or commercially available assay kits based on the Gries...

journal_title：Nature protocols

authors： Nussler AK,Glanemann M,Schirmeier A,Liu L,Nüssler NC

更新日期：2006-01-01 00:00:00

abstract：:By determining protein-protein interactions in normal, diseased and infected cells, we can improve our understanding of cellular systems and their reaction to various perturbations. In this protocol, we discuss how to use data obtained in affinity purification-mass spectrometry (AP-MS) experiments to generate meaningf...

journal_title：Nature protocols

authors： Morris JH,Knudsen GM,Verschueren E,Johnson JR,Cimermancic P,Greninger AL,Pico AR

更新日期：2014-11-01 00:00:00

abstract：:At present, several assays that use radioisotope labeling to quantify the degradation of long-lived proteins have been developed to measure autophagic flux. Here, we describe a nonradioactive pulse-chase protocol using L-azidohomoalanine (AHA) labeling to quantify long-lived protein degradation during autophagy. AHA i...

journal_title：Nature protocols

authors： Wang J,Zhang J,Lee YM,Ng S,Shi Y,Hua ZC,Lin Q,Shen HM

更新日期：2017-12-01 00:00:00

abstract：:Here we describe a protocol for the production of frozen competent yeast cells that can be transformed with high efficiency using the lithium acetate/single-stranded carrier DNA/PEG method. This protocol allows the production of highly competent yeast cells that can be frozen and used at a later date and is especially...

journal_title：Nature protocols

authors： Gietz RD,Schiestl RH

更新日期：2007-01-01 00:00:00

abstract：:Single-cell gene expression analysis has contributed to a better understanding of the transcriptional heterogeneity in a variety of model systems, including those used in research in developmental, cancer and stem cell biology. Nowadays, technological advances facilitate the generation of large gene expression data se...

journal_title：Nature protocols

authors： Durruthy-Durruthy R,Gottlieb A,Heller S

更新日期：2015-03-01 00:00:00

abstract：:Cell-to-cell chemical signaling plays multiple roles in coordinating the activity of the functional elements of an organism, with these elements ranging from a three-neuron reflex circuit to the entire animal. In recent years, single-cell mass spectrometry (MS) has enabled the discovery of cell-to-cell signaling molec...

journal_title：Nature protocols

authors： Rubakhin SS,Sweedler JV

更新日期：2007-01-01 00:00:00

abstract：:The plant cell wall is a chemically complex structure composed mostly of polysaccharides. Detailed analyses of these cell wall polysaccharides are essential for our understanding of plant development and for our use of plant biomass (largely wall material) in the food, agriculture, fabric, timber, biofuel and biocompo...

journal_title：Nature protocols

authors： Pettolino FA,Walsh C,Fincher GB,Bacic A

更新日期：2012-09-01 00:00:00

abstract：:Neurons in cortical sensory regions receive modality-specific information through synapses that are located on their dendrites. Recently, the use of two-photon microscopy combined with whole-cell recordings has helped to identify visually evoked dendritic calcium signals in mouse visual cortical neurons in vivo. The c...

journal_title：Nature protocols

authors： Jia H,Rochefort NL,Chen X,Konnerth A

更新日期：2011-01-01 00:00:00

abstract：:Biominerals serve as critical structures of living systems and play important roles in biochemical processes. Understanding their crystallization mechanisms is therefore central to many areas of biology, biogeoscience, and biochemistry. Some biominerals, such as bone and dentin, are hierarchical nanocomposite structur...

journal_title：Nature protocols

authors： Zhang X,He Y,Liu J,Bowden ME,Kovarik L,Mao SX,Wang C,De Yoreo JJ,Rosso KM

更新日期：2018-09-01 00:00:00

abstract：:This protocol describes a method for directing the expression of genes of interest into postmitotic neocortical neurons in vivo. Microinjection of a DNA plasmid-amphiphilic molecule mix into the neocortex followed by delivery of an ad hoc electric pulse protocol during the first few days of life in mice allows rapid, ...

journal_title：Nature protocols

authors： De la Rossa A,Jabaudon D

更新日期：2015-01-01 00:00:00

abstract：:The small ubiquitin-like modifier (SUMO) is a protein modifier that is post-translationally coupled to thousands of lysines in more than a thousand proteins. An understanding of which lysines are modified by SUMO is critical in unraveling its function as a master regulator of all nuclear processes, as well as its invo...

journal_title：Nature protocols

authors： Hendriks IA,Vertegaal AC

更新日期：2016-09-01 00:00:00

abstract：:Ex vivo perfusion systems offer a reliable, reproducible method for studying acute physiological responses of an organ to various environmental manipulations. Unlike in vitro culture systems, the cellular organization, compartmentalization and three-dimensional structure of ex vivo-perfused organs are maintained. Thes...

journal_title：Nature protocols

authors： Goeden N,Bonnin A

更新日期：2013-01-01 00:00:00

abstract：:Recombineering is an efficient method of in vivo genetic engineering applicable to chromosomal as well as episomal replicons in Escherichia coli. This method circumvents the need for most standard in vitro cloning techniques. Recombineering allows construction of DNA molecules with precise junctions without constraint...

journal_title：Nature protocols

authors： Sharan SK,Thomason LC,Kuznetsov SG,Court DL

更新日期：2009-01-01 00:00:00

abstract：:The human stomach contains two primary domains: the corpus, which contains the fundic epithelium, and the antrum. Each of these domains has distinct cell types and functions, and therefore each presents with unique disease pathologies. Here, we detail two protocols to differentiate human pluripotent stem cells (hPSCs)...

journal_title：Nature protocols

authors： Broda TR,McCracken KW,Wells JM

更新日期：2019-01-01 00:00:00

abstract：:Understanding chemical reactivity through the use of state-of-the-art computational techniques enables chemists to both predict reactivity and rationally design novel reactions. This protocol aims to provide chemists with the tools to implement a powerful and robust method for analyzing and understanding any chemical ...

journal_title：Nature protocols

authors： Vermeeren P,van der Lubbe SCC,Fonseca Guerra C,Bickelhaupt FM,Hamlin TA

更新日期：2020-02-01 00:00:00

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：:Determining enantiomeric excess (e.e.) in chiral compounds is key to development of chiral catalyst auxiliaries and chiral drugs. Here we describe a sensitive and robust fluorescence-based assay for determining e.e. in mixtures of enantiomers of 1,2- and 1,3-diols, chiral amines, amino alcohols, and amino-acid esters....

journal_title：Nature protocols

authors： Shcherbakova EG,James TD,Anzenbacher P Jr

更新日期：2020-07-01 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：:A procedure for the conversion of a symmetrical ketone to an enantiomerically pure lactam is described. The technique described here involves a ring-expansion reaction of a 4-substituted cyclohexanone accomplished with a chiral 1,3-azidopropanol derivative. The procedure entails first a one-step preparation of (R)-1-p...

journal_title：Nature protocols

authors： Ribelin TP,Aubé J

更新日期：2008-01-01 00:00:00

abstract：:The Golgi apparatus undergoes extensive disassembly during mitosis and reassembly in post-mitotic daughter cells. This process has been mimicked in vitro by treating Golgi membranes with mitotic and interphase cytosol. To determine the minimal machinery that controls the morphological change, we have developed a defin...

journal_title：Nature protocols

authors： Tang D,Xiang Y,Wang Y

更新日期：2010-04-01 00:00:00

abstract：:Post-translational modifications alter protein structure, affecting activity, stability, localization and/or binding partners. Antibodies that specifically recognize post-translationally modified proteins have a number of uses including immunocytochemistry and immunoprecipitation of the modified protein to purify prot...

journal_title：Nature protocols

authors： Arur S,Schedl T

更新日期：2014-02-01 00:00:00

abstract：:Long-read sequencing technologies have become increasingly popular due to their strengths in resolving complex genomic regions. As a leading model organism with small genome size and great biotechnological importance, the budding yeast Saccharomyces cerevisiae has many isolates currently being sequenced with long read...

journal_title：Nature protocols

authors： Yue JX,Liti G

更新日期：2018-06-01 00:00:00

abstract：:One of the challenges for modern neuroscience is to understand the rules of concerted neuronal function in vivo. This question can be addressed using noninvasive high-resolution imaging techniques like two-photon microscopy. This protocol describes a versatile approach for in vivo two-photon calcium imaging of neural ...

journal_title：Nature protocols

authors： Garaschuk O,Milos RI,Konnerth A

更新日期：2006-01-01 00:00:00

abstract：:Because RNA-protein interactions have a central role in a wide array of biological processes, methods that enable a quantitative assessment of these interactions in a high-throughput manner are in great demand. Recently, we developed the high-throughput sequencing-RNA affinity profiling (HiTS-RAP) assay that couples s...

journal_title：Nature protocols

authors： Ozer A,Tome JM,Friedman RC,Gheba D,Schroth GP,Lis JT

更新日期：2015-08-01 00:00:00

abstract：:Cell proliferation may be measured in vivo by quantifying DNA synthesis with isotopically labeled deoxyribonucleotide precursors. Deuterium-labeled glucose is one such precursor which, because it achieves high levels of enrichment for a short period, is well suited to the study of rapidly dividing cells, in contrast t...

journal_title：Nature protocols

authors： Macallan DC,Asquith B,Zhang Y,de Lara C,Ghattas H,Defoiche J,Beverley PC

更新日期：2009-01-01 00:00:00

abstract：:As proteins within cells are spatially organized according to their role, knowledge about protein localization gives insight into protein function. Here, we describe the LOPIT technique (localization of organelle proteins by isotope tagging) developed for the simultaneous and confident determination of the steady-stat...

journal_title：Nature protocols

authors： Sadowski PG,Dunkley TP,Shadforth IP,Dupree P,Bessant C,Griffin JL,Lilley KS

更新日期：2006-01-01 00:00:00