A protocol for isolation and culture of human umbilical vein endothelial cells.

abstract：:Postembryonic development is an important process of organismal maturation after embryonic growth. Despite key progress in recent years in understanding embryonic development via fluorescence time-lapse microscopy, comparatively less live-cell imaging of postembryonic development has been done. Here we describe a prot...

journal_title：Nature protocols

authors： Chai Y,Li W,Feng G,Yang Y,Wang X,Ou G

更新日期：2012-12-01 00:00:00

abstract：:This protocol describes the primary culture of individual chromaffin cells derived by enzymatic digestion from the adrenal medulla of the bovine adrenal gland. Since the late 1970s, such cells have provided a useful model system to study neurotransmitter biosynthesis, storage and release in the catecholaminergic syste...

journal_title：Nature protocols

authors： O'Connor DT,Mahata SK,Mahata M,Jiang Q,Hook VY,Taupenot L

更新日期：2007-01-01 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

authors： Nelson CM,Inman JL,Bissell MJ

更新日期：2008-01-01 00:00:00

abstract：:This protocol describes a method combining phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers to characterize the germination of single bacterial spores. The characterization consists of the following steps: (i) loading heat-activated dormant spores into a temperature-controlled micros...

journal_title：Nature protocols

authors： Kong L,Zhang P,Wang G,Yu J,Setlow P,Li YQ

更新日期：2011-05-01 00:00:00

abstract：:Optogenetic tools provide users the ability to photocontrol the activity of cells. Commonly, activation is achieved by expression of proteins from photosynthetic organisms, for example, microbial opsins (e.g., ChR2). Alternatively, a sister approach, synthetic optogenetics, enables photocontrol over proteins of mammal...

journal_title：Nature protocols

authors： Carmi I,De Battista M,Maddalena L,Carroll EC,Kienzler MA,Berlin S

更新日期：2019-03-01 00:00:00

abstract：:We describe a protocol for the insertion of ultrashort single-walled carbon nanotubes (SWCNTs) to form nanopores in a Montal-Mueller lipid bilayer. The SWCNTs are designed to bind to a specific analyte of interest; binding will result in the reduction of current in single-channel recording experiments. The first stage...

journal_title：Nature protocols

authors： Liu L,Xie J,Li T,Wu HC

更新日期：2015-11-01 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：:This transformation procedure generates, with high efficiency (70-90%), hairy roots in cultivars, landraces and accessions of Phaseolus vulgaris (common bean) and other Phaseolus spp. Hairy roots rapidly develop after wounding young plantlets with Agrobacterium rhizogenes, at the cotyledon node, and keeping the plants...

journal_title：Nature protocols

authors： Estrada-Navarrete G,Alvarado-Affantranger X,Olivares JE,Guillén G,Díaz-Camino C,Campos F,Quinto C,Gresshoff PM,Sanchez F

更新日期：2007-01-01 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

authors： Maroney PA,Chamnongpol S,Souret F,Nilsen TW

更新日期：2008-01-01 00:00:00

abstract：:An important area of proteomics involves the need for quantification, whether relative or absolute. Many methods now exist for relative quantification, but to support biomarker proteomics and systems biology, absolute quantification rather than relative quantification is required. Absolute quantification usually invol...

journal_title：Nature protocols

authors： Pratt JM,Simpson DM,Doherty MK,Rivers J,Gaskell SJ,Beynon RJ

更新日期：2006-01-01 00:00:00

abstract：:N-Succinimidyl 3-(di-tert-butyl[(18)F]fluorosilyl)benzoate ([(18)F]SiFB) is a highly reactive prosthetic group for radiolabeling of proteins for use in positron emission tomography (PET). It is similar to N-succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB), the 'gold-standard' prosthetic group for protein (18)F-labeli...

journal_title：Nature protocols

authors： Kostikov AP,Chin J,Orchowski K,Schirrmacher E,Niedermoser S,Jurkschat K,Iovkova-Berends L,Wängler C,Wängler B,Schirrmacher R

更新日期：2012-11-01 00:00:00

abstract：:Pseudo-afferent cervical lymph-duct cannulation in a sheep model allows large amounts of lymph cells to be collected under physiological conditions, carrying immune signaling information from the head tissues, including oro-nasal mucosae. Importantly, large quantities of dendritic cells (DCs) of several subtypes are o...

journal_title：Nature protocols

authors： Schwartz-Cornil I,Epardaud M,Bonneau M

更新日期：2006-01-01 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

authors： Dahl JA,Collas P

更新日期：2008-01-01 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

authors： Wang J,Allgeyer ES,Sirinakis G,Zhang Y,Hu K,Lessard MD,Li Y,Diekmann R,Phillips MA,Dobbie IM,Ries J,Booth MJ,Bewersdorf J

更新日期：2020-12-16 00:00:00

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

authors： Fantin A,Vieira JM,Plein A,Maden CH,Ruhrberg C

更新日期：2013-02-01 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：:This protocol describes an in vitro approach for measuring the kinetics and affinities of interactions between membrane-anchored proteins. This method is particularly established for dissecting the interaction dynamics of cytokines with their receptor subunits. For this purpose, the receptor subunits are tethered in a...

journal_title：Nature protocols

authors： Gavutis M,Lata S,Piehler J

更新日期：2006-01-01 00:00:00

abstract：:Hi-C is a powerful method that provides pairwise information on genomic regions in spatial proximity in the nucleus. Hi-C requires millions of cells as input and, as genome organization varies from cell to cell, a limitation of Hi-C is that it only provides a population average of genome conformations. We developed si...

journal_title：Nature protocols

authors： Nagano T,Lubling Y,Yaffe E,Wingett SW,Dean W,Tanay A,Fraser P

更新日期：2015-12-01 00:00:00

abstract：:We describe a simple and versatile scheme to prepare a series of poly(ethylene glycol)-based bidentate ligands that permit strong interactions with colloidal semiconductor nanocrystals (quantum dots, QDs) and gold nanoparticles (AuNPs) alike and promote their dispersion in aqueous solutions. These ligands are synthesi...

journal_title：Nature protocols

authors： Mei BC,Susumu K,Medintz IL,Mattoussi H

更新日期：2009-01-01 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：:Fast, high-resolution mapping of heterogeneous interfaces with a wide elastic modulus range is a major goal of atomic force microscopy (AFM). This goal becomes more challenging when the nanomechanical mapping involves biomolecules in their native environment. Over the years, several AFM-based methods have been develop...

journal_title：Nature protocols

authors： Benaglia S,Gisbert VG,Perrino AP,Amo CA,Garcia R

更新日期：2018-12-01 00:00:00

abstract：:A fluorescence-based microarray technique that does not require target DNA labeling is detailed. This 'label-free' approach utilizes a cationic, water-soluble conjugated polymer PFBT (poly[9,9'-bis(6''-(N,N,N-trimethylammonium)hexyl)fluorene-co-alt-4,7-(2,1,3-benzothiadiazole) dibromide]), and neutral PNA (peptide nuc...

journal_title：Nature protocols

authors： Sun C,Gaylord BS,Hong JW,Liu B,Bazan GC

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

authors： Slot JW,Geuze HJ

更新日期：2007-01-01 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

authors： Berchowitz LE,Copenhaver GP

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

authors： Lerou PH,Yabuuchi A,Huo H,Miller JD,Boyer LF,Schlaeger TM,Daley GQ

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：: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：:Here, we describe a simple protocol for the design and construction of a laser-guided direct writing (LGDW) system able to micropattern the self-assembly of liver sinusoid-like structures with micrometer resolution in vitro. To the best of our knowledge, LGDW is the only technique able to pattern cells "on the fly" wi...

journal_title：Nature protocols

authors： Nahmias Y,Odde DJ

更新日期：2006-01-01 00:00:00

abstract：:Here, we describe a robust protocol for the reverse transfection of cells on small interfering (siRNA) arrays, which, in combination with multi-channel immunofluorescence or time-lapse microscopy, is suitable for genome-wide RNA interference (RNAi) screens in intact human cells. The automatic production of 48 'transfe...

journal_title：Nature protocols

authors： Erfle H,Neumann B,Liebel U,Rogers P,Held M,Walter T,Ellenberg J,Pepperkok R

更新日期：2007-01-01 00:00:00

abstract：:Here we describe a protocol to dissect the peroxisomal matrix protein import pathway using a cell-free in vitro system. The system relies on a postnuclear supernatant (PNS), which is prepared from rat/mouse liver, to act as a source of peroxisomes and cytosolic components. A typical in vitro assay comprises the follow...

journal_title：Nature protocols

authors： Rodrigues TA,Francisco T,Dias AF,Pedrosa AG,Grou CP,Azevedo JE

更新日期：2016-12-01 00:00:00