Abstract:
:This protocol describes a methodology for imaging the sequestration of infected erythrocytes of the rodent malaria parasite Plasmodium berghei in the bodies of live mice or in dissected organs, using a transgenic parasite that expresses luciferase. Real-time imaging of infected erythrocytes is performed by measuring bioluminescence produced by the enzymatic reaction between luciferase and its substrate luciferin, which is injected into the mice several minutes prior to imaging. The bioluminescence signal is detected by an intensified charge-coupled device (I-CCD) photon-counting video camera. Sequestration of infected erythrocytes is imaged during short-term infections with synchronous parasite development or during ongoing infections. With this technology, sequestration patterns of the schizont stage can be quantitatively analyzed within 1-2 d after infection. Real-time in vivo imaging of infected erythrocytes will provide increased insights into the dynamics of sequestration and its role in pathology, and can be used to evaluate strategies that prevent sequestration.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Franke-Fayard B,Waters AP,Janse CJdoi
10.1038/nprot.2006.69subject
Has Abstractpub_date
2006-01-01 00:00:00pages
476-85issue
1eissn
1754-2189issn
1750-2799pii
nprot.2006.69journal_volume
1pub_type
杂志文章相关文献
Nature Protocols文献大全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
pub_type: 杂志文章
doi:10.1038/nprot.2014.017
更新日期:2014-02-01 00:00:00
abstract::This protocol describes a directed evolution method for in vitro mutagenesis and recombination of polynucleotide sequences. The staggered extension process (StEP) is essentially a modified PCR that uses highly abbreviated annealing and extension steps to generate staggered DNA fragments and promote crossover events al...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.309
更新日期:2006-01-01 00:00:00
abstract::This protocol describes a method for the extraction of DNA from elephant ivory. These techniques are being used to assign geographic origin to poached ivory by comparing the ivory genotype to a geographic-based gene frequency map, developed separately. The method has three components: ivory pulverization, decalcificat...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.318
更新日期:2007-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2011.382
更新日期:2011-09-22 00:00:00
abstract::The scototaxis (dark/light preference) protocol is a behavioral model for fish that is being validated to assess the antianxiety effects of pharmacological agents and the behavioral effects of toxic substances, and to investigate the (epi)genetic bases of anxiety-related behavior. Briefly, a fish is placed in a centra...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.225
更新日期:2010-02-01 00:00:00
abstract::The membrane-permeant fluorogenic biarsenicals FlAsH-EDT(2) and ReAsH-EDT(2) can be prepared in good yields by a straightforward two-step procedure from the inexpensive precursor dyes fluorescein and resorufin, respectively. Handling of toxic reagents such as arsenic trichloride is minimized so the synthesis can be ca...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.144
更新日期:2008-01-01 00:00:00
abstract::Isolated protoplasts serve as a transient expression system that is highly representative of stable transgenics in terms of transcriptome responses. They can also be used as a cellular system to study gene transactivation and nucleocytoplasmic protein trafficking. They are particularly useful for systems studies in wh...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.147
更新日期:2014-09-01 00:00:00
abstract::We present a protocol for visualizing and quantifying single mRNA molecules in mammalian (mouse and human) tissues. In the approach described here, sets of about 50 short oligonucleotides, each labeled with a single fluorophore, are hybridized to target mRNAs in tissue sections. Each set binds to a single mRNA molecul...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.109
更新日期:2013-09-01 00:00:00
abstract::This protocol outlines steps for optimizing the transfection of adherent primary mammalian cells using the readily available off-the-shelf cationic polymer, 25-kDa branched polyethylenimine (bPEI25). Transfection efficiency of cationic polymers varies among cell lines and is highly dependent on the conditions and envi...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2012.038
更新日期:2012-04-19 00:00:00
abstract::This protocol and the accompanying software program called LEVER (lineage editing and validation) enable quantitative automated analysis of phase-contrast time-lapse images of cultured neural stem cells. Images are captured at 5-min intervals over a period of 5-15 d as the cells proliferate and differentiate. LEVER au...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2011.422
更新日期:2011-11-17 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
pub_type: 杂志文章
doi:10.1038/nprot.2016.153
更新日期:2017-01-01 00:00:00
abstract::The ability to rapidly generate large panels of antigen-specific human antibodies in a rodent would enable the efficient discovery of novel therapeutically useful antibodies. We have developed a system wherein human antigen-specific antibody-secreting plasmablasts can be enriched in vivo, in a severe combined immunode...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.104
更新日期:2014-07-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2007.373
更新日期:2007-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2006.76
更新日期:2006-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2006.483
更新日期:2007-01-01 00:00:00
abstract::Chromatin and transcriptional processes are among the most intensively studied fields of biology today. The introduction of chromatin immunoprecipitations (ChIP) represents a major advancement in this area. This powerful method allows researchers to probe specific protein-DNA interactions in vivo and to estimate the d...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.27
更新日期:2006-01-01 00:00:00
abstract::In the version of this protocol originally published, the caption for Fig. 3 was erroneously placed with Fig. 4, and that for Fig. 4 was placed with Fig. 3. This error has been corrected in the HTML and PDF versions of the paper. ...
journal_title:Nature protocols
pub_type: 已发布勘误
doi:10.1038/s41596-018-0079-5
更新日期:2019-07-01 00:00:00
abstract::Multipotent neural crest stem cells (NCSCs) have the potential to generate a wide range of cell types including melanocytes; peripheral neurons; and smooth muscle, bone, cartilage and fat cells. This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent c...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2012.156
更新日期:2013-01-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2009.29
更新日期:2009-01-01 00:00:00
abstract::Because RNA-protein interactions have a central role in a wide array of biological processes, methods that enable a quantitative assessment of these interactions in a high-throughput manner are in great demand. Recently, we developed the high-throughput sequencing-RNA affinity profiling (HiTS-RAP) assay that couples s...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.074
更新日期:2015-08-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2011.367
更新日期:2011-08-11 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
pub_type: 杂志文章
doi:10.1038/s41596-020-00463-4
更新日期:2021-01-27 00:00:00
abstract::This protocol describes a method combining phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers to characterize the germination of single bacterial spores. The characterization consists of the following steps: (i) loading heat-activated dormant spores into a temperature-controlled micros...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2011.307
更新日期:2011-05-01 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
pub_type: 杂志文章
doi:10.1038/s41596-019-0150-x
更新日期:2019-05-01 00:00:00
abstract::We report here a high-throughput method for the modification of bacterial artificial chromosomes (BACs) that uses a novel two-plasmid approach. In this protocol, a vector modified in our laboratory to hold an R6Kγ origin of replication and a marker recombination cassette is inserted into a BAC in a single recombinatio...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2010.131
更新日期:2010-09-01 00:00:00
abstract::A protocol for ester condensation between equimolar amounts of carboxylic acids and alcohols catalyzed by bulky diarylammonium pentafluorobenzenesulfonate is described. We also present procedures for the synthesis of N-(2,6-diisopropylphenyl)-N-mesitylammonium pentafluorobenzenesulfonate. The present ester condensatio...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.254
更新日期:2007-01-01 00:00:00
abstract::The small ubiquitin-like modifier (SUMO) is a protein modifier that is post-translationally coupled to thousands of lysines in more than a thousand proteins. An understanding of which lysines are modified by SUMO is critical in unraveling its function as a master regulator of all nuclear processes, as well as its invo...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.082
更新日期:2016-09-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2009.6
更新日期:2009-01-01 00:00:00
abstract::Methods for installing natural and unnatural amino acids and their modifications into proteins in a benign and precise manner are highly sought-after in protein science. Here we describe a protocol for 'post-translational mutagenesis' that enables the programmed installation of protein side chains through the use of r...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.087
更新日期:2017-10-01 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
pub_type: 杂志文章
doi:10.1038/nprot.2017.159
更新日期:2018-04-01 00:00:00