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::The elucidation of protein-protein interaction networks is a crucial task in the postgenomic era. In this protocol, we describe our approach to discover protein-protein interactions using the surface plasmon resonance technique coupled to mass spectrometry (MS). A peptide or a protein is immobilized on a sensor chip a...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.84
更新日期:2009-01-01 00:00:00
abstract::Many techniques have been developed for studying inducible gene expression, but all of them are multicomponent systems consisting of cis-acting elements at the DNA or RNA level, trans-acting regulator proteins and/or small molecules as inducers. RNA thermometers are the only known single-component regulators of gene e...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.112
更新日期:2009-01-01 00:00:00
abstract::Glycoproteins are involved in diverse biological processes ranging from extracellular contact and recognition to intracellular signaling. Crystal structures of glycoproteins would yield tremendous insight into these processes. But glycoprotein structural analysis has been hindered by difficulties in expressing milligr...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.29
更新日期:2009-01-01 00:00:00
abstract::Reactive oxygen species (ROS) are continuously produced in the cell as a consequence of aerobic metabolism, and are controlled by several antioxidant mechanisms. An accurate measurement of ROS is essential to evaluate the redox status of the cell, or the effects of molecules with the pro-oxidant or antioxidant activit...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.189
更新日期:2009-01-01 00:00:00
abstract::Association studies can focus on candidate gene(s), a particular genomic region, or adopt a genome-wide association approach, each of which has implications for marker selection. The strategy for marker selection will affect the statistical power of the study to detect a disease association and is a crucial element of...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.38
更新日期:2009-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2009.32
更新日期:2009-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2009.100
更新日期:2009-01-01 00:00:00
abstract::The systematic mapping of protein interactions by bait-prey techniques, including affinity purification-mass spectrometry or the yeast two-hybrid system, contributes a unique and relevant perspective on the comprehensive picture of cellular machines. We describe here a protocol for statistical analysis of node-and-edg...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.26
更新日期:2009-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2009.6
更新日期:2009-01-01 00:00:00
abstract::Embryonic stem cells (ESCs) constitute a tool of great potential in neurobiology, enabling the directed differentiation of specific neural cell types. We have shown recently that neurons of the cerebral cortex can be generated from mouse ESCs cultured in a chemically defined medium that contains no morphogen, but in t...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.157
更新日期:2009-01-01 00:00:00
abstract::The effect of genetic mutation on phenotype is of significant interest in genetics. The type of genetic mutation that causes a single amino acid substitution (AAS) in a protein sequence is called a non-synonymous single nucleotide polymorphism (nsSNP). An nsSNP could potentially affect the function of the protein, sub...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.86
更新日期:2009-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2008.46
更新日期:2008-01-01 00:00:00
abstract::Human embryonic stem (hES) cells are self-renewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. Most hES cell lines are derived from cryopreserved human embryos that were created during in vitro fertilization (IVF) and are in excess of clinical need. Embryos th...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.60
更新日期:2008-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2008.158
更新日期:2008-01-01 00:00:00
abstract::This protocol describes the principles and methods used for the preparation of cryopreserved cell stocks. Following these procedures will ensure the availability of reproducible cultures for use within a single laboratory at different times and for different collaborating laboratories. Although the basic principle is ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.190
更新日期:2008-01-01 00:00:00
abstract::This protocol explains how to discover functional signals in genomic sequences by detecting over- or under-represented oligonucleotides (words) or spaced pairs thereof (dyads) with the Regulatory Sequence Analysis Tools (http://rsat.ulb.ac.be/rsat/). Two typical applications are presented: (i) predicting transcription...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.98
更新日期:2008-01-01 00:00:00
abstract::The bimolecular fluorescence complementation (BiFC) assay is a powerful tool for visualizing and identifying protein interactions in living cells. This assay is based on the principle of protein-fragment complementation, using two nonfluorescent fragments derived from fluorescent proteins. When two fragments are broug...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.16
更新日期:2008-01-01 00:00:00
abstract::Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.35
更新日期:2008-01-01 00:00:00
abstract::Hydroxyl radical footprinting has been widely used for studying the structure of DNA and DNA-protein complexes. The high reactivity and lack of base specificity of the hydroxyl radical makes it an excellent probe for high-resolution footprinting of DNA-protein complexes; this technique can provide structural detail th...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.72
更新日期:2008-01-01 00:00:00
abstract::Realizing the potential of human embryonic stem cells (hESCs) in research and commercial applications requires generic protocols for culture, expansion and genetic modification that function between multiple lines. Here we describe a feeder-free hESC culture protocol that was tested in 13 independent hESC lines derive...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.140
更新日期:2008-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2008.144
更新日期:2008-01-01 00:00:00
abstract::Photoconvertible fluorescent proteins, such as Kaede, can be switched irreversibly from their native color to a new one. This property can be exploited to visualize de novo mRNA translation, because newly synthesized proteins can be distinguished from preexisting ones by their color. In this protocol, Kaede cDNA linke...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.113
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
pub_type: 杂志文章
doi:10.1038/nprot.2007.537
更新日期:2008-01-01 00:00:00
abstract::The point mutational spectrum over nearly any 75- to 250-bp DNA sequence isolated from cells, tissues or large populations may be discovered using denaturing capillary electrophoresis (DCE). A modification of the standard DCE method that uses cycling temperature (e.g., +/-5 degrees C), CyDCE, permits optimal resolutio...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.79
更新日期:2008-01-01 00:00:00
abstract::Desorption electrospray ionization (DESI) allows the direct analysis of ordinary objects or pre-processed samples under ambient conditions. Among other applications, DESI is used to identify and record spatial distributions of lipids and drug molecules in biological tissue sections. This technique does not require sam...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.11
更新日期:2008-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2007.491
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
pub_type: 杂志文章
doi:10.1038/nprot.2007.518
更新日期:2008-01-01 00:00:00
abstract::MicroRNAs (miRNAs) are short, about 21 nucleotides in length, noncoding, regulatory RNA molecules representing a new layer in post-transcriptional regulation of gene expression. Intensive miRNA research has necessitated the development of effective miRNA detection methods such as northern analyses, quantitative real-t...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.528
更新日期:2008-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2007.530
更新日期:2008-01-01 00:00:00
abstract::In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) a...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.182
更新日期:2008-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2008.68
更新日期:2008-01-01 00:00:00
abstract::We present a detailed protocol for the structural analysis of protein-linked glycans. In this approach, appropriate for glycomics studies, N-linked glycans are released using peptide N-glycosidase F and O-linked glycans are released by reductive alkaline beta-elimination. Using strategies based on mass spectrometry (m...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.227
更新日期:2007-01-01 00:00:00
abstract::The partial reduction of electron-deficient pyrroles using either Birch (Li/NH(3)) or ammonia-free (Li/di-tert-butyl biphenyl) conditions allows formation of pyrroline compounds in good yield and, when combined with a reductive alkylation or similar approach, leads to highly functionalized, synthetically useful compou...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.245
更新日期:2007-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2007.277
更新日期:2007-01-01 00:00:00
abstract::Studies on colonic cells in the lamina propria (LP) of mice are important for understanding the cellular and immune responses in the gut, especially in inflammatory bowel diseases (such as morbus crohn and colitis ulcerosa). This protocol details a method to isolate LP cells and characterize freshly isolated cells by ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.315
更新日期:2007-01-01 00:00:00
abstract::A detailed protocol for the synthesis of core/shell semiconductor nanocrystal, their encapsulation into phospholipid micelles, their purification and their coupling to a controlled number of small molecules is given. The protocol for the core/shell quantum dot (QD) CdSe/CdZnS synthesis has been specifically designed w...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.351
更新日期:2007-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2007.365
更新日期:2007-01-01 00:00:00
abstract::Protein phosphorylation plays important roles in various aspects of cellular events. Visualization of site-specific phosphorylation in cells is of great importance not only to analyze spatial and temporal distribution but also to investigate biological function. Now, site- and phosphorylation state-specific antibodies...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.374
更新日期:2007-01-01 00:00:00
abstract::Apoptosis plays a pivotal role in the regulation of cell turnover, and a defect or an excess of apoptosis has been implicated in several human diseases. Apoptosis is activated from an extracellular death signal, or from an internal pathway starting from the endoplasmatic reticulum or the mitochondria. To investigate t...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.397
更新日期:2007-01-01 00:00:00
abstract::This protocol presents the peptide incorporation of environment-sensitive fluorophores derived from the dimethylaminophthalimide family. The procedure utilizes anhydride precursors of 4-dimethylaminophthalimide (4-DMAP) or 6-dimethylaminonaphthalimide (6-DMN), whose syntheses are described in a related protocol from t...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.442
更新日期:2007-01-01 00:00:00