A ring barrier-based migration assay to assess cell migration in vitro.

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

authors： Mulvey CM,Breckels LM,Geladaki A,Britovšek NK,Nightingale DJH,Christoforou A,Elzek M,Deery MJ,Gatto L,Lilley KS

更新日期：2017-06-01 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：:The rodent malaria parasite Plasmodium chabaudi chabaudi shares many features with human malaria species, including P. falciparum, and is the in vivo model of choice for many aspects of malaria research in the mammalian host, from sequestration of parasitized erythrocytes, to antigenic variation and host immunity and ...

journal_title：Nature protocols

authors： Spence PJ,Cunningham D,Jarra W,Lawton J,Langhorne J,Thompson J

更新日期：2011-04-01 00:00:00

abstract：:The CRISPR-Cas9 genome-editing tool and the availability of whole-genome sequences from plant species have revolutionized our ability to introduce targeted mutations into important crop plants, both to explore genetic changes and to introduce new functionalities. Here, we describe protocols adapting the CRISPR-Cas9 sy...

journal_title：Nature protocols

authors： Osakabe Y,Liang Z,Ren C,Nishitani C,Osakabe K,Wada M,Komori S,Malnoy M,Velasco R,Poli M,Jung MH,Koo OJ,Viola R,Nagamangala Kanchiswamy C

更新日期：2018-12-01 00:00:00

abstract：:Fluorescent microspheres are commonly used to assess bone blood supply in large animals, but the technique is not widely used in smaller mammals, as traditional methods such as reference blood sampling, ventilation and catheterization are not easily applied. This protocol describes a viable alternative for measuring b...

journal_title：Nature protocols

authors： Serrat MA

更新日期：2009-01-01 00:00:00

abstract：:This protocol describes how to purify and radiolabel collagen for use as a substrate to assay collagenolytic members of the matrix metalloproteinases (MMPs). This assay measures enzymes that specifically cleave native triple helical collagen. After incubation of the MMP enzyme with the collagen substrate at 37 degrees...

journal_title：Nature protocols

authors： Cawston TE

更新日期：2009-01-01 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

authors： Shcherbakova EG,James TD,Anzenbacher P Jr

更新日期：2020-07-01 00:00:00

abstract：:Iterative saturation mutagenesis (ISM) is a new and efficient method for the directed evolution of functional enzymes. It reduces the necessary molecular biological work and the screening effort drastically. It is based on a Cartesian view of the protein structure, performing iterative cycles of saturation mutagenesis...

journal_title：Nature protocols

authors： Reetz MT,Carballeira JD

更新日期：2007-01-01 00:00:00

abstract：:Neurons in cortical sensory regions receive modality-specific information through synapses that are located on their dendrites. Recently, the use of two-photon microscopy combined with whole-cell recordings has helped to identify visually evoked dendritic calcium signals in mouse visual cortical neurons in vivo. The c...

journal_title：Nature protocols

authors： Jia H,Rochefort NL,Chen X,Konnerth A

更新日期：2011-01-01 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

authors： Baudin B,Bruneel A,Bosselut N,Vaubourdolle M

更新日期：2007-01-01 00:00:00

abstract：:Using siRNAs to genetically manipulate immune cells is important to both basic immunological studies and therapeutic applications. However, siRNA delivery is challenging because primary immune cells are often sensitive to the delivery materials and generate immune responses. We have recently developed an amphiphilic d...

journal_title：Nature protocols

authors： Chen J,Ellert-Miklaszewska A,Garofalo S,Dey AK,Tang J,Jiang Y,Clément F,Marche PN,Liu X,Kaminska B,Santoni A,Limatola C,Rossi JJ,Zhou J,Peng L

更新日期：2021-01-01 00:00:00

abstract：:The fabrication of 3D tissues retaining the original functions of tissues/organs in vitro is crucial for optimal tissue engineering and regenerative medicine. The fabrication of 3D tissues also contributes to the establishment of in vitro tissue/organ models for drug screening. Our laboratory has developed a fabricati...

journal_title：Nature protocols

authors： Haraguchi Y,Shimizu T,Sasagawa T,Sekine H,Sakaguchi K,Kikuchi T,Sekine W,Sekiya S,Yamato M,Umezu M,Okano T

更新日期：2012-04-05 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：: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

authors： Zheng JS,Tang S,Qi YK,Wang ZP,Liu L

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

authors： Mar AC,Horner AE,Nilsson SR,Alsiö J,Kent BA,Kim CH,Holmes A,Saksida LM,Bussey TJ

更新日期：2013-10-01 00:00:00

abstract：:Isolation of monoclonal antibodies is an important technique for understanding the specificities and characteristics of antibodies that underlie the humoral immune response to a given antigen. Here we describe a technique for isolating monoclonal antibodies from human peripheral blood mononuclear cells. The protocol i...

journal_title：Nature protocols

authors： Huang J,Doria-Rose NA,Longo NS,Laub L,Lin CL,Turk E,Kang BH,Migueles SA,Bailer RT,Mascola JR,Connors M

更新日期：2013-10-01 00:00:00

abstract：:Protein ADP-ribosylation is a structurally heterogeneous post-translational modification (PTM) that influences the physicochemical and biological properties of the modified protein. ADP-ribosylation of chromatin changes its structural properties, thereby regulating important nuclear functions. A lack of suitable antib...

journal_title：Nature protocols

authors： Bisceglie L,Bartolomei G,Hottiger MO

更新日期：2017-09-01 00:00:00

abstract：:RNA interference (RNAi) is an efficient method for silencing genes in cultured cells. Here we describe a simple RNAi approach for silencing genes in a cell type-specific and tissue-specific way in vivo. The approach, which mimics the means by which naturally occurring 'microRNA's are generated, uses a tissue-specific ...

journal_title：Nature protocols

authors： Rao MK,Wilkinson MF

更新日期：2006-01-01 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

authors： Leung KM,Holt CE

更新日期：2008-01-01 00:00:00

abstract：:We describe the use of a microfabricated cell culture substrate, consisting of a uniform array of closely spaced, vertical, elastomeric microposts, to study the effects of substrate rigidity on cell function. Elastomeric micropost substrates are micromolded from silicon masters comprised of microposts of different hei...

journal_title：Nature protocols

authors： Yang MT,Fu J,Wang YK,Desai RA,Chen CS

更新日期：2011-02-01 00:00:00

abstract：:Traumatic brain injury (TBI) is a major cause of mortality and morbidity. Various attempts have been made to replicate clinical TBI using animal models. The fluid-percussion model (FP) is one of the oldest and most commonly used models of experimentally induced TBI. Both central (CFP) and lateral (LFP) variations of t...

journal_title：Nature protocols

authors： Kabadi SV,Hilton GD,Stoica BA,Zapple DN,Faden AI

更新日期：2010-09-01 00:00:00

abstract：:Mass spectrometry-based phosphoproteomic analysis is a powerful method for gaining a global, unbiased understanding of cellular signaling. Its accuracy and comprehensiveness stands or falls with the quality and choice of the applied phosphopeptide prefractionation strategy. This protocol covers a powerful but simple a...

journal_title：Nature protocols

authors： Zarei M,Sprenger A,Rackiewicz M,Dengjel J

更新日期：2016-01-01 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

authors： Dongari-Bagtzoglou A,Kashleva H

更新日期：2006-01-01 00:00:00

abstract：:Coimmunoprecipitation (co-IP) analysis is a useful method for studying protein-protein interactions. It currently involves electrophoresis and western blotting, which are not optimized for detecting weak and transient interactions. In this protocol we describe an advanced version of co-IP analysis that uses real-time,...

journal_title：Nature protocols

authors： Lee HW,Ryu JY,Yoo J,Choi B,Kim K,Yoon TY

更新日期：2013-10-01 00:00:00

abstract：:Raman spectroscopy can be used to measure the chemical composition of a sample, which can in turn be used to extract biological information. Many materials have characteristic Raman spectra, which means that Raman spectroscopy has proven to be an effective analytical approach in geology, semiconductor, materials and p...

journal_title：Nature protocols

authors： Butler HJ,Ashton L,Bird B,Cinque G,Curtis K,Dorney J,Esmonde-White K,Fullwood NJ,Gardner B,Martin-Hirsch PL,Walsh MJ,McAinsh MR,Stone N,Martin FL

更新日期：2016-04-01 00:00:00

abstract：:Carbohydrate microarrays have received considerable attention as an advanced technology for the rapid analysis of carbohydrate-protein interactions. This protocol provides detailed procedures for the preparation of carbohydrate microarrays by immobilizing hydrazide-conjugated carbohydrates on epoxide-derivatized glass...

journal_title：Nature protocols

authors： Park S,Lee MR,Shin I

更新日期：2007-01-01 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：: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

authors： Ebada SS,Edrada RA,Lin W,Proksch P

更新日期：2008-01-01 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

authors： Goeden N,Bonnin A

更新日期：2013-01-01 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

authors： Carion O,Mahler B,Pons T,Dubertret B

更新日期：2007-01-01 00:00:00