Generation of human antral and fundic gastric organoids from pluripotent stem cells.

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：:In recent years, many mouse models have been developed to mark and trace the fate of adult cell populations using fluorescent proteins. High-resolution visualization of such fluorescent markers in their physiological setting is thus an important aspect of adult stem cell research. Here we describe a protocol to produc...

journal_title：Nature protocols

authors： Snippert HJ,Schepers AG,Delconte G,Siersema PD,Clevers H

更新日期：2011-07-28 00:00:00

abstract：:A mechanistic understanding of HIV-1 latency depends on a model system that recapitulates the in vivo condition of latently infected, resting CD4(+) T lymphocytes. Latency seems to be established after activated CD4(+) T cells, the principal targets of HIV-1 infection, become productively infected and survive long eno...

journal_title：Nature protocols

authors： Kim M,Hosmane NN,Bullen CK,Capoferri A,Yang HC,Siliciano JD,Siliciano RF

更新日期：2014-12-01 00:00:00

abstract：:Here we describe a protocol for the production of frozen competent yeast cells that can be transformed with high efficiency using the lithium acetate/single-stranded carrier DNA/PEG method. This protocol allows the production of highly competent yeast cells that can be frozen and used at a later date and is especially...

journal_title：Nature protocols

authors： Gietz RD,Schiestl RH

更新日期：2007-01-01 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：: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：:By determining protein-protein interactions in normal, diseased and infected cells, we can improve our understanding of cellular systems and their reaction to various perturbations. In this protocol, we discuss how to use data obtained in affinity purification-mass spectrometry (AP-MS) experiments to generate meaningf...

journal_title：Nature protocols

authors： Morris JH,Knudsen GM,Verschueren E,Johnson JR,Cimermancic P,Greninger AL,Pico AR

更新日期：2014-11-01 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

authors： Pettersson FH,Anderson CA,Clarke GM,Barrett JC,Cardon LR,Morris AP,Zondervan KT

更新日期：2009-01-01 00:00:00

abstract：:Accurate analysis of DNA sequence variation in not only humans and animals but also other organisms has played a significant role in expanding our knowledge about genetic variety and diversity in a number of different biological areas. The search for an understanding of the causes of genetic variants and mutations has...

journal_title：Nature protocols

authors： Ota M,Fukushima H,Kulski JK,Inoko H

更新日期：2007-01-01 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：:We describe procedures for the synthesis of a fluorescent pyrimidine analog and its site-specific incorporation into a DNA oligomer. The 5'-protected and 3'-activated nucleoside 4 is synthesized in three steps with an overall yield of 40%. Site-specific incorporation into a DNA oligomer occurs with greater than 88% co...

journal_title：Nature protocols

authors： Greco NJ,Tor Y

更新日期：2007-01-01 00:00:00

abstract：:The protein called 'small ubiquitin-like modifier' (SUMO) is post-translationally linked to target proteins at the ɛ-amino group of lysine residues. This 'SUMOylation' alters the behavior of the target protein, a change that is utilized to regulate diverse cellular processes. Understanding the target-specific conseque...

journal_title：Nature protocols

authors： Tammsalu T,Matic I,Jaffray EG,Ibrahim AF,Tatham MH,Hay RT

更新日期：2015-09-01 00:00:00

abstract：:Surface- and matrix-bound signals modulate stem cell fate in vivo and in vitro. This protocol enables the immobilization of a wide range of biomolecules that contain primary amino groups to different types of solid carriers, including glass substrates and standard polystyrene well plates. We describe how thin polymer ...

journal_title：Nature protocols

authors： Pompe T,Salchert K,Alberti K,Zandstra P,Werner C

更新日期：2010-06-01 00:00:00

abstract：:Proteins are continually produced and degraded, to avoid the accumulation of old or damaged molecules and to maintain the efficiency of physiological processes. Despite its importance, protein turnover has been difficult to measure in vivo. Previous approaches to evaluating turnover in vivo have required custom labeli...

journal_title：Nature protocols

authors： Alevra M,Mandad S,Ischebeck T,Urlaub H,Rizzoli SO,Fornasiero EF

更新日期：2019-12-01 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

authors： Sun N,Lee A,Wu JC

更新日期：2009-01-01 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：: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：:Targeted nucleases, including zinc-finger nucleases (ZFNs), transcription activator-like (TAL) effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9), have provided researchers with the ability to manipulate nearly any genomic sequence in hu...

journal_title：Nature protocols

authors： Liu J,Gaj T,Yang Y,Wang N,Shui S,Kim S,Kanchiswamy CN,Kim JS,Barbas CF 3rd

更新日期：2015-11-01 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：: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：: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：:Here we present a protocol for the extraction of RNA from Dictyostelium discoideum. Dictyostelium is a social amoeba that undergoes a basic developmental program, and therefore analysis of RNA levels over a time course is a commonly used technique. This procedure is similar to other guanidine thiocyanate-based methods...

journal_title：Nature protocols

authors： Pilcher KE,Gaudet P,Fey P,Kowal AS,Chisholm RL

更新日期：2007-01-01 00:00:00

abstract：:Social recognition (SR) enables rodents to distinguish between familiar and novel conspecifics, largely through individual odor cues. SR tasks utilize the tendency for a male to sniff and interact with a novel individual more than a familiar individual. Many paradigms have been used to study the roles of the neuropept...

journal_title：Nature protocols

authors： Macbeth AH,Edds JS,Young WS 3rd

更新日期：2009-01-01 00:00:00

abstract：:This protocol describes a culture system in which inner-ear sensory tissue is produced from mouse embryonic stem (ES) cells under chemically defined conditions. This model is amenable to basic and translational investigations into inner ear biology and regeneration. In this protocol, mouse ES cells are aggregated in 9...

journal_title：Nature protocols

authors： Koehler KR,Hashino E

更新日期：2014-01-01 00:00:00

abstract：:This protocol describes a general method for the preparation of RNAs in which the reactivity or hydrogen-bonding properties of the molecule are modified in a photoreversible fashion by use of a caging strategy. A single caged adenosine, modified at the 2' position as a nitro-benzyl ether, can be incorporated into shor...

journal_title：Nature protocols

authors： Chaulk SG,MacMillan AM

更新日期：2007-01-01 00:00:00

abstract：:Apoptosis is a critical factor in AIDS and other viral illnesses, cerebral and myocardial ischemia, autoimmune and neurodegenerative states, organ and bone marrow transplant rejection, and tumor response to chemotherapy and radiation. Improved methods to identify sites of apoptosis are increasing our understanding of ...

journal_title：Nature protocols

authors： Blankenberg FG,Vanderheyden JL,Strauss HW,Tait JF

更新日期：2006-01-01 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 analysis of large microbiome data sets holds great promise for the delineation of the biological and metabolic functioning of living organisms and their role in the environment. In the midst of this genomic puzzle, viruses, especially those that infect microbial communities, represent a major reservoir of genetic ...

journal_title：Nature protocols

authors： Paez-Espino D,Pavlopoulos GA,Ivanova NN,Kyrpides NC

更新日期：2017-08-01 00:00:00

abstract：:Prediction of protein-protein interactions at the structural level on the proteome scale is important because it allows prediction of protein function, helps drug discovery and takes steps toward genome-wide structural systems biology. We provide a protocol (termed PRISM, protein interactions by structural matching) f...

journal_title：Nature protocols

authors： Tuncbag N,Gursoy A,Nussinov R,Keskin O

更新日期：2011-08-11 00:00:00

abstract：:The analysis of highly hydrophobic proteins is still an analytical challenge. Using a recombinant gamma-aminobutyric acid A (GABAA)-receptor subunit as a model protein, we developed a gel-based proteomic approach for high MS/MS-peptide sequence coverage identification. Protein samples were separated by multi-dimension...

journal_title：Nature protocols

authors： Kang SU,Fuchs K,Sieghart W,Pollak A,Csaszar E,Lubec G

更新日期：2009-01-01 00:00:00