解剖学和形态学
麻醉学
听力与言语-语言病理学
行为科学
心脏和心血管系统
细胞和组织工程学
临床神经病学
危重症监护医学
牙科,口腔外科和医学
皮肤病学
急诊医学
内分泌学和新陈代谢
肠胃学和肝脏学
老人病学和老年医学
卫生保健科学和服务
血液学
免疫学
传染病
综合和补充性医学
医学伦理学
医学信息学
医学实验室技术
医学,全科和内科
医学,法律
医学,研究和试验
神经系统科学
护理
营养学和饮食学
产科医学和妇科医学
肿瘤学
眼科学
整形外科学
耳鼻喉科学
病理学
儿科学
周围血管疾病
药理学和药剂学
生理学
基本医疗保健
精神病学
公共、环境和职业卫生
放射学,核医学和医学成像
康复学
生殖生物学
呼吸系统
风湿病学
运动科学
外科学
毒理学
热带医学
泌尿学和肾脏学
病毒学
老年医学
健康政策和服务
心理学,临床
abstract::Here we describe a protocol for advanced CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis). The CUBIC protocol enables simple and efficient organ clearing, rapid imaging by light-sheet microscopy and quantitative imaging analysis of multiple samples. The organ or body is cleared by im...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.085
更新日期:2015-11-01 00:00:00
abstract::The direct detection of drug-protein interactions in living cells is a major challenge in drug discovery research. Recently, we introduced an approach termed thermal proteome profiling (TPP), which enables the monitoring of changes in protein thermal stability across the proteome using quantitative mass spectrometry. ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.101
更新日期:2015-10-01 00:00:00
abstract::The protein called 'small ubiquitin-like modifier' (SUMO) is post-translationally linked to target proteins at the ɛ-amino group of lysine residues. This 'SUMOylation' alters the behavior of the target protein, a change that is utilized to regulate diverse cellular processes. Understanding the target-specific conseque...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.095
更新日期:2015-09-01 00:00:00
abstract::In order to meaningfully analyze common and rare genetic variants, results from genome-wide association studies (GWASs) of multiple cohorts need to be combined in a meta-analysis in order to obtain enough power. This requires all cohorts to have the same single-nucleotide polymorphisms (SNPs) in their GWASs. To this e...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.077
更新日期:2015-09-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2015.084
更新日期:2015-09-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2015.074
更新日期:2015-08-01 00:00:00
abstract::Cell migration is a key feature of virtually every biological process, and it can be studied in a variety of ways. Here we outline a protocol for the in vitro study of cell migration using a ring barrier-based assay. A 'barrier' is inserted in the culture chamber, which prevents cells from entering a defined area. Cel...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.056
更新日期:2015-06-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2015.048
更新日期:2015-05-01 00:00:00
abstract::AMPylation (adenylylation) has been recognized as an important post-translational modification that is used by pathogens to regulate host cellular proteins and their associated signaling pathways. AMPylation has potential functions in various cellular processes, and it is widely conserved across both prokaryotes and e...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.044
更新日期:2015-05-01 00:00:00
abstract::In this protocol, we describe the preparation of nanoscale iron oxide-based materials and their use in the catalysis of different hydrogenation reactions. Pyrolysis of a Fe(OAc)2-phenanthroline complex on carbon at 800 °C under argon atmosphere results in the formation of nanoscale Fe2O3 particles surrounded by nitrog...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.025
更新日期:2015-04-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2015.030
更新日期:2015-03-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2015.022
更新日期:2015-03-01 00:00:00
abstract::Hepatic stellate cells (HSCs) have been identified as the main fibrogenic cell type in the liver. Hence, efforts to understand hepatic fibrogenesis and to develop treatment strategies have focused on this cell type. HSC isolation, originally developed in rats, has subsequently been adapted to mice, thus allowing the s...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.017
更新日期:2015-02-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2015.018
更新日期:2015-02-01 00:00:00
abstract::Raman and Fourier transform IR (FTIR) microspectroscopic images of biological material (tissue sections) contain detailed information about their chemical composition. The challenge lies in identifying changes in chemical composition, as well as locating and assigning these changes to different conditions (pathology, ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.008
更新日期:2015-02-01 00:00:00
abstract::Small ubiquitin-like modifier (SUMO) post-translational modification (PTM) of proteins has a crucial role in the regulation of important cellular processes. This protocol describes the chemical synthesis of functional SUMO-peptide conjugates. The two crucial stages of this protocol are the solid-phase synthesis of pep...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.013
更新日期:2015-02-01 00:00:00
abstract::Herein we describe a protocol that uses hollow-fiber flow field-flow fractionation (FFF) coupled with multiangle light scattering (MALS) for hydrodynamic size-based separation and characterization of complex protein aggregates. The fractionation method, which requires 1.5 h to run, was successfully modified from the a...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.009
更新日期:2015-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2015.001
更新日期:2015-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2014.188
更新日期:2014-12-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2014.164
更新日期:2014-11-01 00:00:00
abstract::Influenza A viruses (IAVs) cause epidemics and pandemics that result in considerable financial burden and loss of human life. To manage annual IAV epidemics and prepare for future pandemics, an improved understanding of how IAVs emerge, transmit, cause disease and acquire pandemic potential is urgently needed. Fundame...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.180
更新日期:2014-11-01 00:00:00
abstract::In situ mRNA hybridization is one of the most powerful techniques for analyzing patterns of gene expression. However, its usefulness is limited in complex plant tissues by the need to fix, embed and section samples before localizing the desired mRNA. Here we present a robust whole-mount in situ hybridization method th...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.162
更新日期:2014-10-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2014.147
更新日期:2014-09-01 00:00:00
abstract::Overexpression screens can be used to explore gene function in Drosophila melanogaster, but to demonstrate their full potential, comprehensive and systematic collections of fly strains are required. Here we provide a protocol for high-throughput cloning of Drosophila open-reading frames (ORFs) that are regulated by up...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.105
更新日期:2014-07-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2014.104
更新日期:2014-07-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2014.065
更新日期:2014-05-01 00:00:00
abstract::Post-translational protein nitration has attracted interest owing to its involvement in cellular signaling, effects on protein function and potential as biomarker of nitroxidative stress. We describe a procedure for enriching nitropeptides for mass spectrometry (MS)-based proteomics that is a simple and reliable alter...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.052
更新日期:2014-04-01 00:00:00
abstract::The pig has emerged as an important large animal model in biomedical and pharmaceutical research. We describe a protocol for high-efficiency germline transgenesis and sustained transgene expression in pigs by using the Sleeping Beauty (SB) transposon system. The protocol is based on co-injection of a plasmid encoding ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.010
更新日期:2014-04-01 00:00:00
abstract::The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen-independent microfluidic CTC-...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.044
更新日期:2014-03-01 00:00:00
abstract::Mouse embryonic stem cells (mESCs) are key tools for genetic engineering, development of stem cell-based therapies and basic research on pluripotency and early lineage commitment. However, successful derivation of germline-competent embryonic stem cell lines has, until recently, been limited to a small number of inbre...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.030
更新日期:2014-03-01 00:00:00
abstract::Dynamic interplay between intracellular organelles requires a particular functional apposition of membrane structures. The organelles involved come into close contact, but do not fuse, thereby giving rise to notable microdomains; these microdomains allow rapid communication between the organelles. Plasma membrane-asso...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.016
更新日期:2014-02-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2014.017
更新日期:2014-02-01 00:00:00
abstract::This protocol describes a culture system in which inner-ear sensory tissue is produced from mouse embryonic stem (ES) cells under chemically defined conditions. This model is amenable to basic and translational investigations into inner ear biology and regeneration. In this protocol, mouse ES cells are aggregated in 9...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.100
更新日期:2014-01-01 00:00:00
abstract::Characterization of the cellular participants in tissue immune responses is crucial to understanding infection, cancer, autoimmunity, allergy, graft rejection and other immunological processes. Previous reports indicate that leukocytes in lung vasculature fail to be completely removed by perfusion. Several studies sug...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.005
更新日期:2014-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2013.163
更新日期:2014-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2013.173
更新日期:2014-01-01 00:00:00
abstract::The combination of ChIP-seq and transcriptome analysis is a compelling approach to unravel the regulation of gene expression. Several recently published methods combine transcription factor (TF) binding and gene expression for target prediction, but few of them provide an efficient software package for the community. ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.150
更新日期:2013-12-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2013.152
更新日期:2013-12-01 00:00:00
abstract::We recently invented a method to produce highly potent siRNAs in Escherichia coli, based on the serendipitous discovery that ectopic expression of p19, a plant viral siRNA-binding protein, stabilizes otherwise unstable bacterial siRNAs, which we named pro-siRNAs for prokaryotic siRNAs. We present a detailed protocol d...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.149
更新日期:2013-12-01 00:00:00
abstract::This protocol details a subset of assays developed within the touchscreen platform to measure various aspects of executive function in rodents. Three main procedures are included: extinction, measuring the rate and extent of curtailing a response that was previously, but is no longer, associated with reward; reversal ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.123
更新日期:2013-10-01 00:00:00