Site-specific protein labeling using PRIME and chelation-assisted click chemistry.

abstract：:This protocol describes the synthesis of oligo(poly(ethylene glycol) fumarate) (OPF; 1-35 kDa; a polymer useful for tissue engineering applications) by a one-pot reaction of poly(ethylene glycol) (PEG) and fumaryl chloride. The procedure involves three parts: dichloromethane and PEG are first dried; the reaction step ...

journal_title：Nature protocols

authors： Kinard LA,Kasper FK,Mikos AG

更新日期：2012-05-31 00:00:00

abstract：:Spectroscopic techniques such as Fourier-transform infrared (FTIR) spectroscopy are used to study interactions of light with biological materials. This interaction forms the basis of many analytical assays used in disease screening/diagnosis, microbiological studies, and forensic/environmental investigations. Advantag...

journal_title：Nature protocols

authors： Morais CLM,Paraskevaidi M,Cui L,Fullwood NJ,Isabelle M,Lima KMG,Martin-Hirsch PL,Sreedhar H,Trevisan J,Walsh MJ,Zhang D,Zhu YG,Martin FL

更新日期：2019-05-01 00:00:00

abstract：:Adult stem cell-based organoid technology is a versatile tool for the generation and long-term maintenance of near-native 3D epithelial tissues in vitro. The generation of cancer organoids from primary patient material enables a range of therapeutic agents to be tested in the resulting organoid cultures. Patient-deriv...

journal_title：Nature protocols

authors： Driehuis E,Kretzschmar K,Clevers H

更新日期：2020-10-01 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：: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：:Hepatic stellate cells (HSCs) have been identified as the main fibrogenic cell type in the liver. Hence, efforts to understand hepatic fibrogenesis and to develop treatment strategies have focused on this cell type. HSC isolation, originally developed in rats, has subsequently been adapted to mice, thus allowing the s...

journal_title：Nature protocols

authors： Mederacke I,Dapito DH,Affò S,Uchinami H,Schwabe RF

更新日期：2015-02-01 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：: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：: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：:Unlike transplantation of other solid organs, vascularized mouse lung transplantation has only recently been developed. In this protocol, we describe a detailed method for performing a vascularized and aerated mouse orthotopic lung transplant, which to date represents the most physiological mouse model of lung transpl...

journal_title：Nature protocols

authors： Krupnick AS,Lin X,Li W,Okazaki M,Lai J,Sugimoto S,Richardson SB,Kornfeld CG,Garbow JR,Patterson GA,Gelman AE,Kreisel D

更新日期：2009-01-01 00:00:00

abstract：:Post-translational modifications alter protein structure, affecting activity, stability, localization and/or binding partners. Antibodies that specifically recognize post-translationally modified proteins have a number of uses including immunocytochemistry and immunoprecipitation of the modified protein to purify prot...

journal_title：Nature protocols

authors： Arur S,Schedl T

更新日期：2014-02-01 00:00:00

abstract：:A big challenge in proteomics is the identification of cell-type-specific proteomes in vivo. This protocol describes how to label, purify and identify cell-type-specific proteomes in living mice. To make this possible, we created a Cre-recombinase-inducible mouse line expressing a mutant methionyl-tRNA synthetase (L27...

journal_title：Nature protocols

authors： Alvarez-Castelao B,Schanzenbächer CT,Langer JD,Schuman EM

更新日期：2019-02-01 00:00:00

abstract：:The direct detection of drug-protein interactions in living cells is a major challenge in drug discovery research. Recently, we introduced an approach termed thermal proteome profiling (TPP), which enables the monitoring of changes in protein thermal stability across the proteome using quantitative mass spectrometry. ...

journal_title：Nature protocols

authors： Franken H,Mathieson T,Childs D,Sweetman GM,Werner T,Tögel I,Doce C,Gade S,Bantscheff M,Drewes G,Reinhard FB,Huber W,Savitski MM

更新日期：2015-10-01 00:00:00

abstract：:Dynamic interplay between intracellular organelles requires a particular functional apposition of membrane structures. The organelles involved come into close contact, but do not fuse, thereby giving rise to notable microdomains; these microdomains allow rapid communication between the organelles. Plasma membrane-asso...

journal_title：Nature protocols

authors： Suski JM,Lebiedzinska M,Wojtala A,Duszynski J,Giorgi C,Pinton P,Wieckowski MR

更新日期：2014-02-01 00:00:00

abstract：:The telomeric repeat amplification protocol (TRAP) is a two-step process for analyzing telomerase activity in cell or tissue extracts. Recent modifications of this sensitive assay include elimination of radioactivity by using a fluorescently labeled primer instead of a radiolabeled primer. In addition, the TRAP assay ...

journal_title：Nature protocols

authors： Herbert BS,Hochreiter AE,Wright WE,Shay JW

更新日期：2006-01-01 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：:Mouse embryonic stem cells (mESCs) are key tools for genetic engineering, development of stem cell-based therapies and basic research on pluripotency and early lineage commitment. However, successful derivation of germline-competent embryonic stem cell lines has, until recently, been limited to a small number of inbre...

journal_title：Nature protocols

authors： Czechanski A,Byers C,Greenstein I,Schrode N,Donahue LR,Hadjantonakis AK,Reinholdt LG

更新日期：2014-03-01 00:00:00

abstract：:Limbal stem cells (LSCs) have an important role in the maintenance of the corneal surface epithelium, and autologous cultured limbal epithelial cell transplantations have contributed substantially to the treatment of the visually disabling condition known as LSC deficiency. In this protocol, we describe a method of es...

journal_title：Nature protocols

authors： Mariappan I,Maddileti S,Savy S,Tiwari S,Gaddipati S,Fatima A,Sangwan VS,Balasubramanian D,Vemuganti GK

更新日期：2010-08-01 00:00:00

abstract：:Rapid evaluation of microbial cell-surface carbohydrates is essential to understanding the mechanisms by which bacteria use glycans to establish pathogenic or symbiotic relationships. Microbial glycan analysis is complicated both by the vast diversity of possible carbohydrate structures and by their dynamic nature. Ba...

journal_title：Nature protocols

authors： Hsu KL,Mahal LK

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：:Biominerals serve as critical structures of living systems and play important roles in biochemical processes. Understanding their crystallization mechanisms is therefore central to many areas of biology, biogeoscience, and biochemistry. Some biominerals, such as bone and dentin, are hierarchical nanocomposite structur...

journal_title：Nature protocols

authors： Zhang X,He Y,Liu J,Bowden ME,Kovarik L,Mao SX,Wang C,De Yoreo JJ,Rosso KM

更新日期：2018-09-01 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：:Although differences in protein staining intensity can often be visualized by difference gel electrophoresis, abundant proteins can obscure less abundant proteins, and quantification of post-translational modifications is difficult. We present a protocol for quantifying changes in the abundance of a specific protein o...

journal_title：Nature protocols

authors： Asara JM,Zhang X,Zheng B,Maroney LA,Christofk HR,Wu N,Cantley LC

更新日期：2006-01-01 00:00:00

abstract：:The research community is in a race to understand the molecular mechanisms of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, to repurpose currently available antiviral drugs and to develop new therapies and vaccines against coronavirus disease 2019 (COVID-19). One major challenge in achieving ...

journal_title：Nature protocols

authors： Gurumurthy CB,Quadros RM,Richardson GP,Poluektova LY,Mansour SL,Ohtsuka M

更新日期：2020-12-01 00:00:00

abstract：:Cell-to-cell chemical signaling plays multiple roles in coordinating the activity of the functional elements of an organism, with these elements ranging from a three-neuron reflex circuit to the entire animal. In recent years, single-cell mass spectrometry (MS) has enabled the discovery of cell-to-cell signaling molec...

journal_title：Nature protocols

authors： Rubakhin SS,Sweedler JV

更新日期：2007-01-01 00:00:00

abstract：:Activity-based protein profiling (ABPP) has emerged as a valuable chemical proteomics method to guide the therapeutic development of covalent drugs by assessing their on-target engagement and off-target activity. We recently used ABPP to determine the serine hydrolase interaction landscape of the experimental drug BIA...

journal_title：Nature protocols

authors： van Rooden EJ,Florea BI,Deng H,Baggelaar MP,van Esbroeck ACM,Zhou J,Overkleeft HS,van der Stelt M

更新日期：2018-04-01 00:00:00

abstract：:Single-cell electroporation allows transfection of plasmid DNA or macromolecules into individual living cells using modified patch electrodes and common electrophysiological equipment. This protocol is optimized for rapid in vivo electroporation of Xenopus laevis tadpole brains with DNA, dextrans, morpholinos and comb...

journal_title：Nature protocols

authors： Bestman JE,Ewald RC,Chiu SL,Cline HT

更新日期：2006-01-01 00:00:00

abstract：:Multiplex-fluorescence in situ hybridization (M-FISH) was initially developed to stain human chromosomes--the 22 autosomes and X and Y sex chromosomes--with uniquely distinctive colors to facilitate karyotyping. The characteristic spectral signatures of all different combinations of fluorochromes are determined by mul...

journal_title：Nature protocols

authors： Geigl JB,Uhrig S,Speicher MR

更新日期：2006-01-01 00:00:00

abstract：:Analysis of the sequence and nature of protein N termini has many applications. Defining the termini of proteins for proteome annotation in the Human Proteome Project is of increasing importance. Terminomics analysis of protease cleavage sites in degradomics for substrate discovery is a key new application. Here we de...

journal_title：Nature protocols

authors： Kleifeld O,Doucet A,Prudova A,auf dem Keller U,Gioia M,Kizhakkedathu JN,Overall CM

更新日期：2011-09-22 00:00:00

abstract：:This protocol outlines the carboxyfluorescein diacetate succinimidyl ester (CFSE) method for following the proliferation of human lymphocytes in vitro and mouse lymphocytes both in vitro and in vivo. The method relies on the ability of CFSE to covalently label long-lived intracellular molecules with the highly fluores...

journal_title：Nature protocols

authors： Quah BJ,Warren HS,Parish CR

更新日期：2007-01-01 00:00:00