Tissue-specific and cell type-specific RNA interference in vivo.

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：:In the version of this protocol originally published, the caption for Fig. 3 was erroneously placed with Fig. 4, and that for Fig. 4 was placed with Fig. 3. This error has been corrected in the HTML and PDF versions of the paper. ...

journal_title：Nature protocols

authors： Cruz-Acuña R,Quirós M,Huang S,Siuda D,Spence JR,Nusrat A,García AJ

更新日期：2019-07-01 00:00:00

abstract：:Venous access catheters used in clinics are prone to biofilm contamination, contributing to chronic and nosocomial infections. Although several animal models for studying device-associated biofilms were previously described, only a few detailed protocols are currently available. Here we provide a protocol using totall...

journal_title：Nature protocols

authors： Chauhan A,Ghigo JM,Beloin C

更新日期：2016-03-01 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：:Isolated protoplasts serve as a transient expression system that is highly representative of stable transgenics in terms of transcriptome responses. They can also be used as a cellular system to study gene transactivation and nucleocytoplasmic protein trafficking. They are particularly useful for systems studies in wh...

journal_title：Nature protocols

authors： Lin YC,Li W,Chen H,Li Q,Sun YH,Shi R,Lin CY,Wang JP,Chen HC,Chuang L,Qu GZ,Sederoff RR,Chiang VL

更新日期：2014-09-01 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：:Unlike humans, mouse bone marrow-derived mesenchymal stem cells (MSCs) cannot be easily harvested by adherence to plastic owing to the contamination of cultures by hematopoietic cells. The design of the protocol described here is based on the phenomenon that compact bones abound in MSCs and hematopoietic cells exist i...

journal_title：Nature protocols

authors： Zhu H,Guo ZK,Jiang XX,Li H,Wang XY,Yao HY,Zhang Y,Mao N

更新日期：2010-03-01 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：: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：: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：: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：: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：: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：: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：:In the past decade, Drosophila has emerged as an ideal model organism for studying the genetic components of sleep as well as its regulation and functions. In fruit flies, sleep can be conveniently estimated by measuring the locomotor activity of the flies using techniques and instruments adapted from the field of cir...

journal_title：Nature protocols

authors： Gilestro GF

更新日期：2012-04-26 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：:Mass spectrometry imaging (MSI) enables label-free spatial mapping of hundreds of biomolecules in tissue sections. This capability provides valuable information on tissue heterogeneity that is difficult to obtain using population-averaged assays. Despite substantial developments in both instrumentation and methodology...

journal_title：Nature protocols

authors： Yin R,Burnum-Johnson KE,Sun X,Dey SK,Laskin J

更新日期：2019-12-01 00:00:00

abstract：:The analysis of large microbiome data sets holds great promise for the delineation of the biological and metabolic functioning of living organisms and their role in the environment. In the midst of this genomic puzzle, viruses, especially those that infect microbial communities, represent a major reservoir of genetic ...

journal_title：Nature protocols

authors： Paez-Espino D,Pavlopoulos GA,Ivanova NN,Kyrpides NC

更新日期：2017-08-01 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：: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：:The membrane-permeant fluorogenic biarsenicals FlAsH-EDT(2) and ReAsH-EDT(2) can be prepared in good yields by a straightforward two-step procedure from the inexpensive precursor dyes fluorescein and resorufin, respectively. Handling of toxic reagents such as arsenic trichloride is minimized so the synthesis can be ca...

journal_title：Nature protocols

authors： Adams SR,Tsien RY

更新日期：2008-01-01 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：:In cell affinity chromatography, type-specific cell separation is based on the interaction between cell-surface receptors and an immobilized ligand on a stationary matrix. This protocol describes the preparation of monolithic polyacrylamide and polydimethylacrylamide cryogel affinity matrices that can be used as a gen...

journal_title：Nature protocols

authors： Kumar A,Srivastava A

更新日期：2010-11-01 00:00:00

abstract：:Development of non-invasive and accurate methods to track cell fate after delivery will greatly expedite transition of embryonic stem (ES) cell therapy to the clinic. In this protocol, we describe the in vivo monitoring of stem cell survival, proliferation and migration using reporter genes. We established stable ES c...

journal_title：Nature protocols

authors： Sun N,Lee A,Wu JC

更新日期：2009-01-01 00:00:00

abstract：:Among the different developed solid-state nanopores, nanopores constructed in a monolayer of molybdenum disulfide (MoS2) stand out as powerful devices for single-molecule analysis or osmotic power generation. Because the ionic current through a nanopore is inversely proportional to the thickness of the pore, ultrathin...

journal_title：Nature protocols

authors： Graf M,Lihter M,Thakur M,Georgiou V,Topolancik J,Ilic BR,Liu K,Feng J,Astier Y,Radenovic A

更新日期：2019-04-01 00:00:00

abstract：:This protocol describes a method to obtain spatially resolved proteomic maps of specific compartments within living mammalian cells. An engineered peroxidase, APEX2, is genetically targeted to a cellular region of interest. Upon the addition of hydrogen peroxide for 1 min to cells preloaded with a biotin-phenol substr...

journal_title：Nature protocols

authors： Hung V,Udeshi ND,Lam SS,Loh KH,Cox KJ,Pedram K,Carr SA,Ting AY

更新日期：2016-03-01 00:00:00

abstract：:Positron emission tomography (PET) is a sensitive and noninvasive imaging method that is widely used to explore molecular events in living subjects. PET can precisely and quantitatively evaluate cellular apoptosis, which has a crucial role in various physiological and pathological processes. In this protocol, we descr...

journal_title：Nature protocols

authors： Wang Z,Wang F,Hida N,Kiesewetter DO,Tian J,Niu G,Chen X

更新日期：2015-05-01 00:00:00

abstract：:The study of angiogenic endothelial cells (ECs) has in recent years greatly stimulated multiple fields of vascular biology research. A number of cellular models and numerous complex developmental, manipulatory and tumor animal models have been developed to study angiogenesis in vitro and in vivo. To connect the versat...

journal_title：Nature protocols

authors： Laib AM,Bartol A,Alajati A,Korff T,Weber H,Augustin HG

更新日期：2009-01-01 00:00:00

abstract：:This protocol measures externalization of aminophospholipids (APLs) to the outside of the plasma membrane using mass spectrometry (MS). APL externalization occurs in numerous events, and it is relevant for transplant medicine, immunity and cancer. In this protocol, externalized APLs are chemically modified by using a ...

journal_title：Nature protocols

authors： Thomas CP,Clark SR,Hammond VJ,Aldrovandi M,Collins PW,O'Donnell VB

更新日期：2014-01-01 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