Immobilization of growth factors on solid supports for the modulation of stem cell fate.

abstract：:The lung epithelium is derived from the endodermal germ layer, which undergoes a complex series of endoderm-mesoderm-mediated signaling events to generate the final arborized network of conducting airways (bronchi, bronchioles) and gas-exchanging units (alveoli). These stages include endoderm induction, anterior-poste...

journal_title：Nature protocols

authors： Miller AJ,Dye BR,Ferrer-Torres D,Hill DR,Overeem AW,Shea LD,Spence JR

更新日期：2019-02-01 00:00:00

abstract：:Mitophagy is a cellular process that selectively removes damaged, old or dysfunctional mitochondria. Defective mitophagy is thought to contribute to normal aging and to various neurodegenerative and cardiovascular diseases. Previous methods used to detect mitophagy in vivo were cumbersome, insensitive and difficult to...

journal_title：Nature protocols

authors： Sun N,Malide D,Liu J,Rovira II,Combs CA,Finkel T

更新日期：2017-08-01 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：:Among the different developed solid-state nanopores, nanopores constructed in a monolayer of molybdenum disulfide (MoS2) stand out as powerful devices for single-molecule analysis or osmotic power generation. Because the ionic current through a nanopore is inversely proportional to the thickness of the pore, ultrathin...

journal_title：Nature protocols

authors： Graf M,Lihter M,Thakur M,Georgiou V,Topolancik J,Ilic BR,Liu K,Feng J,Astier Y,Radenovic A

更新日期：2019-04-01 00:00:00

abstract：:Generation of precisely patterned neural cells from human pluripotent stem cells (hPSCs) is instrumental in developing disease models and stem cell therapies. Here, we provide a detailed 16-d protocol for obtaining high-purity ventral midbrain (VM) dopamine (DA) progenitors for intracerebral transplantation into anima...

journal_title：Nature protocols

authors： Nolbrant S,Heuer A,Parmar M,Kirkeby A

更新日期：2017-09-01 00:00:00

abstract：:Improvement of cellular uptake and cellular localization is still one of the main obstacles to the development of antisense-antigene therapeutics, including peptide nucleic acid (PNA). Cell-penetrating peptides (CPPs) such as Tat peptide and polyarginine have been widely used to improve the cellular uptake of PNA and ...

journal_title：Nature protocols

authors： Shiraishi T,Nielsen PE

更新日期：2006-01-01 00:00:00

abstract：:The glycocalyx comprises glycosylated proteins and lipids and fcorms the outermost layer of cells. It is involved in fundamental inter- and intracellular processes, including non-self-cell and self-cell recognition, cell signaling, cellular structure maintenance, and immune protection. Characterization of the glycocal...

journal_title：Nature protocols

authors： Li Q,Xie Y,Wong M,Barboza M,Lebrilla CB

更新日期：2020-08-01 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

authors： Anderson KG,Mayer-Barber K,Sung H,Beura L,James BR,Taylor JJ,Qunaj L,Griffith TS,Vezys V,Barber DL,Masopust D

更新日期：2014-01-01 00:00:00

abstract：:More than 400,000 people each year suffer adverse effects following bites from venomous snakes. However, snake venom is also a rich source of bioactive molecules with known or potential therapeutic applications. Manually 'milking' snakes is the most common method to obtain venom. Safer alternative methods to produce v...

journal_title：Nature protocols

authors： Puschhof J,Post Y,Beumer J,Kerkkamp HM,Bittenbinder M,Vonk FJ,Casewell NR,Richardson MK,Clevers H

更新日期：2021-01-27 00:00:00

abstract：:The colorimetric bio-barcode assay is a red-to-blue color change-based protein detection method with ultrahigh sensitivity. This assay is based on both the bio-barcode amplification method that allows for detecting miniscule amount of targets with attomolar sensitivity and gold nanoparticle-based colorimetric DNA dete...

journal_title：Nature protocols

authors： Nam JM,Jang KJ,Groves JT

更新日期：2007-01-01 00:00:00

abstract：:Telomeres have emerged as crucial cellular elements in aging and various diseases including cancer. To measure the average length of telomere repeats in cells, we describe our protocols that use fluorescent in situ hybridization (FISH) with labeled peptide nucleic acid (PNA) probes specific for telomere repeats in com...

journal_title：Nature protocols

authors： Baerlocher GM,Vulto I,de Jong G,Lansdorp PM

更新日期：2006-01-01 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

authors： Shinohara H,Wang F,Hossain SM

更新日期：2008-01-01 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：:Conventional gene expression studies analyze multiple cells simultaneously or single cells, for which the exact in vivo or in situ position is unknown. Although cellular heterogeneity can be discerned when analyzing single cells, any spatially defined attributes that underpin the heterogeneous nature of the cells cann...

journal_title：Nature protocols

authors： Chen J,Suo S,Tam PP,Han JJ,Peng G,Jing N

更新日期：2017-03-01 00:00:00

abstract：:The world is on the cusp of a post-antibiotic era, but researchers and medical doctors have found a way forward-by looking back at how infections were treated before the advent of antibiotics, namely using phage therapy. Although bacteriophages (phages) continue to lack drug approval in Western medicine, an increasing...

journal_title：Nature protocols

authors： Luong T,Salabarria AC,Edwards RA,Roach DR

更新日期：2020-09-01 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

authors： Macallan DC,Asquith B,Zhang Y,de Lara C,Ghattas H,Defoiche J,Beverley PC

更新日期：2009-01-01 00:00:00

abstract：:The pollen tube is an excellent single-cell model system for studying cellular processes in plant cell biology. This protocol describes a detailed step-by-step procedure with optimized conditions for introducing various fluorescent reporter proteins into lily, tobacco and Arabidopsis pollen grains by means of biolisti...

journal_title：Nature protocols

authors： Wang H,Jiang L

更新日期：2011-04-01 00:00:00

abstract：:High throughput microarray transcription analyses provide us with the expression profiles for large amounts of plant genes. However, their tissue and cellular resolution is limited. Thus, for detailed functional analysis, it is still necessary to examine the expression pattern of selected candidate genes at a cellular...

journal_title：Nature protocols

authors： Hejátko J,Blilou I,Brewer PB,Friml J,Scheres B,Benková E

更新日期：2006-01-01 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

authors： Sainlos M,Imperiali B

更新日期：2007-01-01 00:00:00

abstract：:Human pluripotent stem cells (hPSCs) provide a valuable model for the study of human development and a means to generate a scalable source of cells for therapeutic applications. This protocol specifies cell fate efficiently into cardiac and endothelial lineages from hPSCs. The protocol takes 2 weeks to complete and re...

journal_title：Nature protocols

authors： Palpant NJ,Pabon L,Friedman CE,Roberts M,Hadland B,Zaunbrecher RJ,Bernstein I,Zheng Y,Murry CE

更新日期：2017-01-01 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

authors： Cato MH,Yau IW,Rickert RC

更新日期：2011-06-09 00:00:00

abstract：:This protocol describes a method for converting short single-stranded and double-stranded DNA into libraries compatible with high-throughput sequencing using Illumina technology. This method has primarily been developed to improve sequence retrieval from ancient DNA, but it is also applicable to the sequencing of shor...

journal_title：Nature protocols

authors： Gansauge MT,Meyer M

更新日期：2013-04-01 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

authors： Das AM,Eggermont AM,ten Hagen TL

更新日期：2015-06-01 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

authors： McDaniel JR,DeKosky BJ,Tanno H,Ellington AD,Georgiou G

更新日期：2016-03-01 00:00:00

abstract：:The small ubiquitin-like modifiers (SUMOs) are posttranslationally conjugated to eukaryotic cellular proteins with generally unpredictable consequences. SUMO substrates are found in many cellular systems, and functional analysis has revealed that substrate SUMOylation often has an important role in their regulation. H...

journal_title：Nature protocols

authors： Tatham MH,Rodriguez MS,Xirodimas DP,Hay RT

更新日期：2009-01-01 00:00:00

abstract：:The Strep-tag II is an eight-residue minimal peptide sequence (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) that exhibits intrinsic affinity toward streptavidin and can be fused to recombinant proteins in various fashions. We describe a protocol that enables quick and mild purification of corresponding Strep-tag II fusion protein...

journal_title：Nature protocols

authors： Schmidt TG,Skerra A

更新日期：2007-01-01 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

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

更新日期：2008-01-01 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

authors： Abuknesha RA,Jeganathan F,Wu J,Baalawy Z

更新日期：2009-01-01 00:00:00

abstract：:To uncover the function of and interplay between the mammalian cytosine modifications 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC), new techniques and advances in current technology are needed. To this end, we have developed oxidative bisulfite sequencing (oxBS-seq), which can quantitatively locate 5mC an...

journal_title：Nature protocols

authors： Booth MJ,Ost TW,Beraldi D,Bell NM,Branco MR,Reik W,Balasubramanian S

更新日期：2013-10-01 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