Enhanced delivery of cell-penetrating peptide-peptide nucleic acid conjugates by endosomal disruption.

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：: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：: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：: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：:Glycoproteins are involved in diverse biological processes ranging from extracellular contact and recognition to intracellular signaling. Crystal structures of glycoproteins would yield tremendous insight into these processes. But glycoprotein structural analysis has been hindered by difficulties in expressing milligr...

journal_title：Nature protocols

authors： Lee JE,Fusco ML,Saphire EO

更新日期：2009-01-01 00:00:00

abstract：:The synthesis of an azobenzene amino acid (aa) for use as a photo-inducible conformational switch in polypeptides is described. The compound can be easily incorporated into an aa sequence by solid-phase peptide synthesis using standard 9-fluorenylmethoxycarbonyl methods. A reversible conformational change of the pepti...

journal_title：Nature protocols

authors： Aemissegger A,Hilvert D

更新日期：2007-01-01 00:00:00

abstract：:Desorption electrospray ionization (DESI) allows the direct analysis of ordinary objects or pre-processed samples under ambient conditions. Among other applications, DESI is used to identify and record spatial distributions of lipids and drug molecules in biological tissue sections. This technique does not require sam...

journal_title：Nature protocols

authors： Wiseman JM,Ifa DR,Venter A,Cooks RG

更新日期：2008-01-01 00:00:00

abstract：:A procedure for the synthesis of N-succinimidyl 3-iodobenzoate labeled with any iodine isotope ([*I]SIB), which is an agent used in the radioiodination of proteins and peptides, from its tin precursor N-succinimidyl 3-(tri-n-butylstannyl)benzoate (STB) is described. Also included are protocols for the synthesis of an ...

journal_title：Nature protocols

authors： Vaidyanathan G,Zalutsky MR

更新日期：2006-01-01 00:00:00

abstract：:The specialized microfold cells (M cells) in the follicle-associated epithelium (FAE) of intestinal Peyer's patches serve as antigen-sampling cells of the intestinal innate immune system. Unlike 'classical' enterocytes, they are able to translocate diverse particulates without digesting them. They act as pathways for ...

journal_title：Nature protocols

authors： Beloqui A,Brayden DJ,Artursson P,Préat V,des Rieux A

更新日期：2017-07-01 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：:This protocol describes an optimized method for direct in vitro monitoring of homo- and heterotypic axon-axon interactions involved in the developmental assembly of neural circuits. The assay exploits a classical example of heterotypic axonal interactions by modeling the sequential extension of spinal motor and somato...

journal_title：Nature protocols

authors： Wang L,Marquardt T

更新日期：2012-01-26 00:00:00

abstract：:Large-insert BAC (bacterial artificial chromosome) and BIBAC (binary BAC) libraries are essential for modern genomics research for all organisms. We helped pioneer the BAC and BIBAC technologies, and by using them we have constructed hundreds of BAC and BIBAC libraries for different species of plants, animals, marine ...

journal_title：Nature protocols

authors： Zhang HB,Scheuring CF,Zhang M,Zhang Y,Wu CC,Dong JJ,Li Y

更新日期：2012-02-16 00:00:00

abstract：:High-throughput ballistic injection nanorheology is a method for the quantitative study of cell mechanics. Cell mechanics are measured by ballistic injection of submicron particles into the cytoplasm of living cells and tracking the spontaneous displacement of the particles at high spatial resolution. The trajectories...

journal_title：Nature protocols

authors： Wu PH,Hale CM,Chen WC,Lee JS,Tseng Y,Wirtz D

更新日期：2012-01-05 00:00:00

abstract：:The immune system operates at the scale of the whole organism in mammals. We currently lack experimental approaches to systematically track and study organism-wide molecular processes in mice. Here we describe an integrated toolkit for measuring gene expression in whole tissues, 3-prime mRNA extension sequencing, that...

journal_title：Nature protocols

authors： Pandey S,Takahama M,Gruenbaum A,Zewde M,Cheronis K,Chevrier N

更新日期：2020-04-01 00:00:00

abstract：:This protocol is an extension to: Nat. Protoc. 6, 870-895 (2014); doi:10.1038/nprot.2011.328; published online 02 June 2011Cycling cells duplicate their DNA content during S phase, following a defined program called replication timing (RT). Early- and late-replicating regions differ in terms of mutation rates, transcr...

journal_title：Nature protocols

authors： Marchal C,Sasaki T,Vera D,Wilson K,Sima J,Rivera-Mulia JC,Trevilla-García C,Nogues C,Nafie E,Gilbert DM

更新日期：2018-05-01 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：: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

authors： Wang S,Sun H,Ma J,Zang C,Wang C,Wang J,Tang Q,Meyer CA,Zhang Y,Liu XS

更新日期：2013-12-01 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：:As proteins within cells are spatially organized according to their role, knowledge about protein localization gives insight into protein function. Here, we describe the LOPIT technique (localization of organelle proteins by isotope tagging) developed for the simultaneous and confident determination of the steady-stat...

journal_title：Nature protocols

authors： Sadowski PG,Dunkley TP,Shadforth IP,Dupree P,Bessant C,Griffin JL,Lilley KS

更新日期：2006-01-01 00:00:00

abstract：:ErmineJ is software for the analysis of functionally interesting patterns in large gene lists drawn from gene expression profiling data or other high-throughput genomics studies. It can be used by biologists with no bioinformatics background to conduct sophisticated analyses of gene sets with multiple methods. It allo...

journal_title：Nature protocols

authors： Gillis J,Mistry M,Pavlidis P

更新日期：2010-06-01 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：:Protein ubiquitination is a versatile protein modification that regulates virtually all cellular processes. This versatility originates from polyubiquitin chains, which can be linked in eight distinct ways. The combinatorial complexity of eight linkage types in homotypic (one chain type per polymer) and heterotypic (m...

journal_title：Nature protocols

authors： Hospenthal MK,Mevissen TET,Komander D

更新日期：2015-02-01 00:00:00

abstract：:Protein digestion using a dedicated protease represents a key element in a typical mass spectrometry (MS)-based shotgun proteomics experiment. Up to now, digestion has been predominantly performed with trypsin, mainly because of its high specificity, widespread availability and ease of use. Lately, it has become appar...

journal_title：Nature protocols

authors： Giansanti P,Tsiatsiani L,Low TY,Heck AJ

更新日期：2016-05-01 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：:Periodontal disease (PD) is a common dental disease associated with the interaction between dysbiotic oral microbiota and host immunity. It is a prevalent disease, resulting in loss of gingival tissue, periodontal ligament, cementum and alveolar bone. PD is a major form of tooth loss in the adult population. Experimen...

journal_title：Nature protocols

authors： Marchesan J,Girnary MS,Jing L,Miao MZ,Zhang S,Sun L,Morelli T,Schoenfisch MH,Inohara N,Offenbacher S,Jiao Y

更新日期：2018-10-01 00:00:00

abstract：:The transmembrane (TM) anchors of cell surface proteins have been one of the 'blind spots' in structural biology because they are generally very hydrophobic, sometimes dynamic, and thus difficult targets for structural characterization. A plethora of examples show these membrane anchors are not merely anchors but can ...

journal_title：Nature protocols

authors： Fu Q,Piai A,Chen W,Xia K,Chou JJ

更新日期：2019-08-01 00:00:00

abstract：:Post-translational protein nitration has attracted interest owing to its involvement in cellular signaling, effects on protein function and potential as biomarker of nitroxidative stress. We describe a procedure for enriching nitropeptides for mass spectrometry (MS)-based proteomics that is a simple and reliable alter...

journal_title：Nature protocols

authors： Prokai L,Guo J,Prokai-Tatrai K

更新日期：2014-04-01 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：: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：: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