Abstract:
:This protocol describes the efficient, generally applicable Ullmann coupling reaction of bromaminic acid with alkyl- or aryl-amines in phosphate buffer under microwave irradiation using elemental copper as a catalyst. The reaction leads to a number of biologically active compounds. As a prototypical example, the synthesis of a new, potent antagonist of human platelet P2Y(12) receptors, which has potential as an antithrombotic drug, is described in detail. The optimized protocol includes a description of an appropriate reaction setup, thin layer chromatography for monitoring the reaction and a procedure for the isolation, purification and characterization of the anticipated product. The reaction is performed without the use of a glove box and there is no requirement for an inert atmosphere. The reaction typically proceeds within 2-30 min, the protocol, including workup, generally takes 1-3 h to complete.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Baqi Y,Müller CEdoi
10.1038/nprot.2010.63subject
Has Abstractpub_date
2010-05-01 00:00:00pages
945-53issue
5eissn
1754-2189issn
1750-2799pii
nprot.2010.63journal_volume
5pub_type
杂志文章相关文献
Nature Protocols文献大全abstract::The mouse embryo hindbrain is a robust and adaptable model for studying sprouting angiogenesis. It permits the spatiotemporal analysis of organ vascularization in normal mice and in mouse strains with genetic mutations that result in late embryonic or perinatal lethality. Unlike postnatal models such as retinal angiog...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.015
更新日期:2013-02-01 00:00:00
abstract::Cap-analysis gene expression (CAGE) provides accurate high-throughput measurement of RNA expression. CAGE allows mapping of all the initiation sites of both capped coding and noncoding RNAs. In addition, transcriptional start sites within promoters are characterized at single-nucleotide resolution. The latter allows t...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2012.005
更新日期:2012-02-23 00:00:00
abstract::Methods for installing natural and unnatural amino acids and their modifications into proteins in a benign and precise manner are highly sought-after in protein science. Here we describe a protocol for 'post-translational mutagenesis' that enables the programmed installation of protein side chains through the use of r...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.087
更新日期:2017-10-01 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::The genomic instability of stem cells in culture, caused by their routine in vitro propagation or by their genetic manipulation, is deleterious both for their clinical application and for their use in basic research. Frequent evaluation of the genomic integrity of stem cells is thus required, and it is usually perform...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.051
更新日期:2013-05-01 00:00:00
abstract::This protocol describes a method for the extraction of DNA from elephant ivory. These techniques are being used to assign geographic origin to poached ivory by comparing the ivory genotype to a geographic-based gene frequency map, developed separately. The method has three components: ivory pulverization, decalcificat...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.318
更新日期:2007-01-01 00:00:00
abstract::The terminal monosaccharide of cell surface glycoconjugates is typically a sialic acid (SA), and aberrant sialylation is involved in several diseases. Several methodological approaches in sample preparation and subsequent analysis using mass spectrometry (MS) have enabled the identification of glycosylation sites and ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2010.167
更新日期:2010-12-01 00:00:00
abstract::Multiplex-fluorescence in situ hybridization (M-FISH) was initially developed to stain human chromosomes--the 22 autosomes and X and Y sex chromosomes--with uniquely distinctive colors to facilitate karyotyping. The characteristic spectral signatures of all different combinations of fluorochromes are determined by mul...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.160
更新日期:2006-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2008.227
更新日期:2009-01-01 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::In this report we describe the development of a standardized three-dimensional (3D) system of the human oral mucosa based on an immortalized human oral keratinocyte cell line (OKF6/TERT-2). The procedure takes approximately 2-3 weeks to complete and includes three main stages: preparation of collagen-embedded fibrobla...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.323
更新日期:2006-01-01 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 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
pub_type: 杂志文章
doi:10.1038/nprot.2017.026
更新日期:2017-06-01 00:00:00
abstract::High-throughput sequencing of the variable domains of immune receptors (antibodies and T cell receptors (TCRs)) is of key importance in the understanding of adaptive immune responses in health and disease. However, the sequencing of both immune receptor chains (VH+VL or TCRβ/δ+TCRα/γ) at the single-cell level for typi...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.024
更新日期:2016-03-01 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
pub_type: 杂志文章
doi:10.1038/s41596-020-0329-1
更新日期:2020-07-01 00:00:00
abstract::The development of single-molecule switching (SMS) fluorescence microscopy (also called single-molecule localization microscopy) over the last decade has enabled researchers to image cell biological structures at unprecedented resolution. Using two opposing objectives in a so-called 4Pi geometry doubles the available ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-020-00428-7
更新日期:2020-12-16 00:00:00
abstract::Gram-negative and Gram-positive bacteria release a variety of membrane vesicles through different formation routes. Knowledge of the structure, molecular cargo and function of bacterial extracellular vesicles (BEVs) is primarily obtained from bacteria cultured in laboratory conditions. BEVs in human body fluids have b...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0236-5
更新日期:2020-01-01 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::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::We report here a high-throughput method for the modification of bacterial artificial chromosomes (BACs) that uses a novel two-plasmid approach. In this protocol, a vector modified in our laboratory to hold an R6Kγ origin of replication and a marker recombination cassette is inserted into a BAC in a single recombinatio...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2010.131
更新日期:2010-09-01 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::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
pub_type: 杂志文章
doi:10.1038/nprot.2012.144
更新日期:2013-01-01 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::Detailed biochemical analysis of unmanipulated germinal center (GC) B cells has not been achieved. Previously, we designed and used a simple, economical and new magnetic bead separation scheme for the purification of 'untouched' mature GC and non-GC B cells from the spleens of immunized mice and reported the first bio...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2011.344
更新日期:2011-06-09 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
abstract::PatternLab for proteomics is an integrated computational environment that unifies several previously published modules for the analysis of shotgun proteomic data. The contained modules allow for formatting of sequence databases, peptide spectrum matching, statistical filtering and data organization, extracting quantit...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.133
更新日期:2016-01-01 00:00:00
abstract::Activity-based protein profiling (ABPP) has emerged as a valuable chemical proteomics method to guide the therapeutic development of covalent drugs by assessing their on-target engagement and off-target activity. We recently used ABPP to determine the serine hydrolase interaction landscape of the experimental drug BIA...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.159
更新日期:2018-04-01 00:00:00
abstract::We describe a protocol for easy isolation and culture of human umbilical vein endothelial cells (HUVECs) to supply every researcher with a method that can be applied in cell biology laboratories with minimum equipment. Endothelial cells (ECs) are isolated from umbilical vein vascular wall by a collagenase treatment, t...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.54
更新日期:2007-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2009.117
更新日期:2009-01-01 00:00:00
abstract::This protocol outlines steps for optimizing the transfection of adherent primary mammalian cells using the readily available off-the-shelf cationic polymer, 25-kDa branched polyethylenimine (bPEI25). Transfection efficiency of cationic polymers varies among cell lines and is highly dependent on the conditions and envi...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2012.038
更新日期:2012-04-19 00:00:00