Mass isotope labeling. Medical search. Frequent questions

Proteins from both cell populations are combined and analyzed together by mass spectrometry as pairs of chemically identical peptides of different stable-isotope composition can be differentiated in a mass spectrometer owing to their mass difference. (wikipedia.org)
The ratio of peak intensities in the mass spectrum for such peptide pairs reflects the abundance ratio for the two proteins. (wikipedia.org)
Here, we describe a new method (immuno-SILAC) for the absolute quantification of proteins in complex samples based on polyclonal antibodies and stable isotope-labeled recombinant protein fragments to allow affinity enrichment prior to mass spectrometry analysis and accurate quantification. (nih.gov)
We identified 231 proteins released from actomyosin contraction-dependent blebs and apoptotic bodies by adapted SILAC (stable isotope labeling with amino acids in cell culture) combined with mass spectrometry analysis. (nature.com)
We used consecutive orthogonal separation platforms to ensure sensitive detection: (a) ion-exchange of intact proteins, (b) SDS-PAGE separation of ion-exchange fractions and (c) HPLC separation of tryptic digests coupled to electrospray tandem mass spectrometry. (hindawi.com)
Isobaric labeling uses stable isotope tags attached to peptides of interest to enable relative or absolute quantitation of proteins via tandem mass spectrometry. (genomeweb.com)
To achieve results of highest possible quality we employed SILAC (Stable Isotope Labeling of Amino acids in Cell culture) to allow quantification, TAP (Tandem Affinity Purification) for recovery, high accuracy Orbitrap mass spectrometry for identification and MaxQuant software for quantification and analysis of SUMO-2 substrate proteins. (europa.eu)
Much of our work involves the use of stable isotope tracers to label proteins in vivo and study synthesis and accumulation of individual proteins. (mayo.edu)
Consequently, a lot of effort is put into identifying the constituent proteins using mass spectrometric methods. (lu.se)

This methodology enables detection of putative metabolites from biological samples and simultaneous quantification of the pattern and extent of isotope labeling. (acs.org)
Absolute quantification with mass spectrometry and isotope labelled internal standards has found broad applications in biomedical research. (nist.gov)
By using our newly developed liquid chromatography-tandem mass spectrometry quantification method, we were able to measure the absolute amount of MRP2/Mrp2 in liver tissues and isolated hepatocytes across species. (aspetjournals.org)
Quantification of Stable Isotope Label in Metabolites via Mass Spectrometry. (mpg.de)

Stable Isotope Labeling by/with Amino acids in Cell culture (SILAC) is a technique based on mass spectrometry that detects differences in protein abundance among samples using non-radioactive isotopic labeling. (wikipedia.org)
A SILAC approach involving incorporation of tyrosine labeled with nine carbon-13 atoms (13C) instead of the normal carbon-12 (12C) has been utilized to study tyrosine kinase substrates in signaling pathways. (wikipedia.org)
Pulsed SILAC (pSILAC) is a variation of the SILAC method where the labelled amino acids are added to the growth medium for only a short period of time. (wikipedia.org)
It had also been used to study biofilm tolerance to antibiotics to differentiate tolerant and sensitive subpopulations Traditionally the level of multiplexing in SILAC was limited due to the number of SILAC isotopes available. (wikipedia.org)
Recently, a new technique called NeuCode (neutron encoding) SILAC, has augmented the level of multiplexing achievable with metabolic labeling (up to 4). (wikipedia.org)
The NeuCode amino acid method is similar to SILAC but differs in that the labeling only utilizes heavy amino acids. (wikipedia.org)
Contains experimentally-derived protein half-life data obtained using stable isotope labeling by amino acids (SILAC) coupled with mass spectrometry. (yeastgenome.org)
According to Josh Coon, professor of chemistry and biomolecular chemistry at UW-Madison and leader of the effort, the new method brings together benefits of SILAC labeling and isobaric tagging. (genomeweb.com)
The method, Coon noted, synthesizes some of the best aspects of isobaric labeling and SILAC, combining, in particular, the multiplexing capabilities of the former and the MS1 level quantitation of the latter. (genomeweb.com)
SILAC - or stable isotope labeling by amino acids in cell culture - uses the incorporation of amino acids containing different isotopes to label proteomic samples for quantitative mass spec analysis. (genomeweb.com)

A protocol based on the simultaneous multiplex capture of more than 40 protein targets showed that approximately half of the antibodies enriched at least one functional peptide detected in the subsequent mass spectrometry analysis. (nih.gov)
Currently standards must be generated one by one by incorporating isotope-labeled amino acids into the peptide sequence by chemical synthesis or by addition to culture media. (genengnews.com)
Digested peptides are labeled with tags that fragment during MS2 to produce signals corresponding to the amount of peptide present in a sample. (genomeweb.com)
However, incorporation of different isotopes significantly increases sample complexity, which can reduce peptide IDs if taken too far. (genomeweb.com)
We describe a statistical measure, Mass Distance Fingerprint, for automatic de novo detection of predominant peptide mass distances, i.e., putative protein mod- ifications. (lu.se)
The method's focus is to globally detect mass differences, not to assign peptide sequences or modifications to individual spectra. (lu.se)
The Mass Distance Fin- gerprint is calculated from high accuracy measured peptide masses. (lu.se)

The combination of immuno-based methods and mass spectrometry detection has great potential in the field of quantitative proteomics. (nih.gov)
Based on recent developments in quantitative mass spectrometry, biological reference materials have been generated that can be used as internal (quality) controls for proteomics utilizing the characteristics of isotope-dilution mass spectrometry (ID-MS). (genengnews.com)
University of Wisconsin-Madison researchers have developed a new labeling approach that could enable significantly higher multiplexing in mass spec proteomics experiments. (genomeweb.com)
Last year, two groups - one led by Harvard University researcher Steven Gygi and another by researchers from chemical proteomics firm Cellzome (since acquired by GlaxoSmithKline) - applied this phenomenon to isobaric tagging, swapping the tags' 12 C atoms for 13 C atoms and 15 N for 14 N to produce a 6-mDa mass shift that upped the reagents' multiplexing capability from six-plex to eight-plex. (genomeweb.com)
In their study released this week, Coon and his team built on this work by incorporating a number of different isotope combinations into lysine molecules, which they then used to metabolically label proteomics samples. (genomeweb.com)
In proteomics, high throughput approaches using mass spectrometry have be- come widely used. (lu.se)

This method uses isotope dilution with carbon-13 labeled internal standards. (cdc.gov)
For assessing the folate content in a range of Brazilian PANCs, we extended the validation of an existing stable isotope dilution assay (SIDA) for the stably 13 C-labelled 10-formyl-Pte[ 13 C 5 ]Glu (10-CHO-Pte[ 13 C 5 ]Glu). (frontiersin.org)
News from the world: The use of D3-Creatine Dilution for muscle mass assessment and its relationship with fracture risk in men. (ectsoc.org)
The deuterated creatine (D 3 Cr) dilution is a novel and interesting methodology that estimates total body creatinine for assessing the overall muscle mass. (ectsoc.org)
the D 3 Cr dilution methodology is employed to assess muscle mass and explore its relationship with incident fragility fractures in a cohort of 1,363 older men from the MrOs study in a mean follow-up time of roughly 4 years and a half. (ectsoc.org)
Interestingly, fractured men had lower muscle mass as assessed by the D 3 Cr dilution methodology, and the lower was the D 3 Cr muscle mass quartile, the higher was the fracture incidence. (ectsoc.org)
Title : Quantitation of trans-fatty acids in human blood via isotope dilution-gas chromatography-negative chemical ionization-mass spectrometry Personal Author(s) : Kuiper, Heather C.;Wei, Na;McGunigale, Samantha L.;Vesper, Hubert W. (cdc.gov)

The combination of high-resolution LC-MS-based untargeted metabolomics with stable isotope tracing provides a global overview of the cellular fate of precursor metabolites. (acs.org)
Recent advances in metabolomics have extended the capability of isotope-assisted studies to reveal global metabolism. (mdpi.com)
For instance, isotope-assisted metabolomics technology has enabled the mapping of a global metabolic network, estimation of flux at branch points of metabolic pathways, and assignment of elemental formulas to unknown metabolites. (mdpi.com)
A reliable method for metabolite extraction is central to mass spectrometry-based metabolomics. (frontiersin.org)

We established an efficient approach, crosslinking of segmentally isotope-labeled RNA and tandem mass spectrometry (CLIR-MS/MS), to localize protein-RNA interactions simultaneously at amino acid and nucleotide resolution. (nih.gov)
Isobaric mass labeling leads to isobaric analytes that can be differentiated by isotopic mass reporters released from the analyte during tandem mass spec. (genengnews.com)
The Tandem Mass Tag® (TMT®) core structure invented by Proteome Sciences allows for the generation of different sets of isotopic and isobaric mass tags. (genengnews.com)
This method is a quantitative procedure for the measurement of nitrate, perchlorate, and thiocyanate in human urine using ion chromatography coupled with electrospray tandem mass spectrometry. (cdc.gov)
These reagents, which are sold as tandem mass tags, or TMT, by Thermo Fisher Scientific through a licensing agreement with Proteome Sciences, and as isobaric tags for relative and absolute quantitation, or iTRAQ, by AB Sciex, enable researchers to multiplex up to eight samples. (genomeweb.com)
Hence, using liquid chromatography-tandem mass spec. (researchgate.net)

The increased multiplexing capability of NeuCode amino acids is from the use of mass defects from extra neutrons in the stable isotopes. (wikipedia.org)

Labeling of Trypanosoma brucei cell cultures with 50% uniformly 13C-labeled glucose demonstrated incorporation of glucose-derived carbon into 187 of 588 putatively identified metabolites in diverse pathways including carbohydrate, nucleotide, lipid, and amino acid metabolism. (acs.org)
Labeling patterns confirmed the metabolic pathways responsible for the biosynthesis of many detected metabolites, and labeling was detected in unexpected metabolites, including two higher sugar phosphates annotated as octulose phosphate and nonulose phosphate. (acs.org)
Multiplexed Profiling and Data Processing Methods to Identify Temperature-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry. (mpg.de)
Analytical platforms such as mass spectrometry and nuclear magnetic resonance (NMR) have emerged as the dominant platforms enabling the robust detection of numerous metabolites. (metabolon.com)
The most comprehensive metabolite coverage is typically accomplished by leveraging a non-targeted approach using high-performance liquid chromatography (HPLC) coupled with high-resolution accurate mass (HRAM) mass spectrometry, commonly referred to as HPLC-MS, or LC-MS. LC-MS is capable of routine detection of thousands of unique chemical compounds, providing broad coverage of biologically relevant metabolites. (metabolon.com)
They compared the isotope patterns of these spectra to the spectra of the metabolites produced from unlabeled glucose and were able to identify herpetopanone, a diterpene with the formula C 20 H 36 O 4 . (chemistryviews.org)
In addition to NMR-based labelling studies, molecular and biochemical tools are used to study the biosynthesis of specialized plant metabolites. (universiteitleiden.nl)

Amyloid β concentrations and stable isotope labeling kinetics of human plasma specific to central nervous system amyloidosis. (msdmanuals.com)

These correspond to a complex mixture of unidentified adducts, 4,5 in-source fragments, multimers, true molecular ions and a vast assortment of isotopes only differentiable by retention time. (metabolon.com)

Markus Nett, Technical University Dortmund, Germany, and colleagues have discovered a new natural product, herpetopanone (pictured), which is produced by Herpetosiphon aurantiacus 114-95 T . The team fed glucose, labeled with 13 C atoms, to a culture of the bacterium, and used liquid chromatography-mass spectrometry (LC-MS) to analyze the metabolic products of the bacteria. (chemistryviews.org)
Analytical methods such as mass spectrometry and nuclear magnetic resonance spectroscopy allow our team to study metabolic consequences of altered energy and protein metabolism. (mayo.edu)

For example, the medium can contain arginine labeled with six carbon-13 atoms (13C) instead of the normal carbon-12 (12C). (wikipedia.org)
Ions from each analyte and each carbon-13 labeled internal standard are monitored, and the abundances of each ion are measured. (cdc.gov)
Using different isotopes of carbon, hydrogen, nitrogen, and oxygen, the researchers worked out 39 distinct isotopologues for lysine spanning a mass range of roughly 39 mDa and separated by 1 mDa each. (genomeweb.com)
It is essential to gain not only information about relative carbon balance and fluxes (common for partitioning studies), but also in absolute mass units. (uni-bayreuth.de)
To evaluate absolute fluxes in belowground carbon pools, eddy covariance and stable isotope labeling were coupled in a 21 day experiment in summer 2010. (uni-bayreuth.de)
Resultant CO 2 assimilation served as absolute carbon input into the ecosystem that was further partitioned by applying the relative carbon distribution in subsidiary pools, gained by 13C pulse labeling and tracing. (uni-bayreuth.de)
Completing carbon isotope sampling campaign in Oslo region. (lu.se)

In general, two different methods of chemical labeling are used-isotopic mass labeling and isobaric mass labeling. (genengnews.com)

Chemical isotope labelling LC-MS analysis methods have been developed for most other phytohormones, enhancing their LC separations and quantitative sensitivity. (unboundmedicine.com)
However, there are currently no reports for chemical-labelled CKs. (unboundmedicine.com)
Here, we report a new chemical isotope labelling LC-MS analytical method for one-pot derivatization of CK bases and their glycoconjugates, based on differential benzylation labelling of the adenine skeleton of CKs with benzyl bromide and its deuterium isotope-labelled reagent. (unboundmedicine.com)
Conversely, chemical labeling, attaching small tags that carry isotope differences to the analytes, presents a simpler approach without the need to actually synthesize any peptides. (genengnews.com)
The method can be used for automated detection of chemical/post- translational modifications, quality control of experiments and labelling approaches, and to control the modification settings of protein identification tools. (lu.se)
The most comprehensive collection of chemical and biological modi- fications being relevant to mass spectrometry can be found in UniMod [12]. (lu.se)

The approach was further developed to also generate quantitative data via the addition of heavy isotope-labeled recombinant protein fragment standards prior to trypsin digestion. (nih.gov)
The kit includes matched, deuterium-labeled internal standards for a total of 44 solutions. (cdc.gov)
Innotope offers "heavy" nucleotides and nucleosides with naturally occurring modifications as mass spectrometry standards, focusing on heavy epigenetic modifiers. (innotope.at)
13 C isotope fully labelled mycotoxins are internal standards for mass spectrometry. (romerlabs.com)
With 13 C fully labelled mycotoxin standards, recovery losses from sample preparation and matrix effects in the MS source can be eliminated. (romerlabs.com)

These small mass differences however need to be resolved on high resolution mass spectrometers. (wikipedia.org)
Detailed in a paper published this week in Nature Methods , the technique, called NeuCode, uses differences in the nuclear binding energy of different isotopes to label amino acids. (genomeweb.com)
Nuclear binding energy is the energy needed to break a nucleus up into its component nucleons, and because every isotope has a unique nuclear binding energy, these differences can be used to distinguish between them. (genomeweb.com)
For the data sets used in this study, known mass differences are detected at electron mass ac- curacy or better. (lu.se)

The eluent from the column is ionized using an electrospray interface to generate and transmit negative ions into the mass spectrometer. (cdc.gov)

Isotopic mass labeling leads to a distinct mass difference of structurally identical reference and target analyte. (genengnews.com)
Comparison of relative response factors (ratio of native analyte to stable isotope labeled internal standard) with known standard concentrations yields individual analyte concentrations. (cdc.gov)

Hoxworth Blood Center will conduct pre-clinical and clinical trials to compare Biotinylation to Chromium-51 to evaluate the safety and efficacy of using a non-radioactive alternative labeling method to assess blood products. (uc.edu)
Under the agreement, BARDA will provide funding and expertise for a clinical trial to validate biotin, a naturally occurring vitamin, as a safe and effective option to replace chromium 51 (Cr-51) as the U.S. Food and Drug Administration's (FDA's) current "gold standard" labeling method for measurement of circulating red blood cells (RBCs). (uc.edu)
Commonly used spectrometric techniques are nuclear magnetic resonance (NMR) and mass spectrometry (MS). Each method has its own pros and cons, while MS prevails in sensitivity and selectivity ( Emwas, 2015 ). (frontiersin.org)

The approach "compresses all the [isotopic] information into tens of milliDaltons, so it's only revealed when you crank up the [mass spec's] resolution," Coon said. (genomeweb.com)

Mass spectrometry-based protein assays impart increased specificity and more rapid development times versus traditional methods, such as ELISA. (brighttalk.com)
These investments rely on enabling technologies, such as safer labeling methods that enable evaluation of circulating RBCs, to support regulatory approval. (uc.edu)
Methods to analyze laser-microdissected samples by NMR spectroscopy and mass spectrometry have been developed in Bernd Schneider's research group and employed for cell type-specific micrometabolic profiling. (universiteitleiden.nl)
2018 ). The later, using similar immunoprecipitation mass spectrometry -based methods, reported that plasma Aβ42 derived measurements can predict brain amyloidosis (measured by amyloid PET) with close to 90% accuracy. (msdmanuals.com)

Their tiny size, rapid growth by clonal propagation, and facile uptake of labeled compounds from the media were attractive features that made them a well-known model for plant biology from 1950 to 1990. (osti.gov)
Advanced wet-lab techniques like isotope-label tracing or measuring mass-to-charge ratios of fragments of compounds are well-established techniques in chemistry. (dagstuhl.de)

The results suggest that polyclonal antibodies generated via immunization of recombinant protein fragments could be used for the enrichment of target peptides to allow for rapid mass spectrometry analysis taking advantage of a substantial reduction in sample complexity. (nih.gov)
The procedure involves enzymatic hydrolysis of urine, extraction, derivatization and analysis using capillary gas chromatog-raphy combined with high resolution mass spectrometry (GC-HRMS). (cdc.gov)
Mass spectral search and analysis using the Golm Metabolome Database. (mpg.de)
Because mass spec resolution isn't infinite, however, the UW-Madison team maxed out at six-plex analysis, running the experiment at 240,000 resolution on a Thermo Fisher Scientific Orbitrap Elite. (genomeweb.com)
Mass [10] together with information on modifications induced by sample prepa- ration procedures for mass spectrometric analysis, but mass changes are only given as integer values. (lu.se)

The researchers found that lower eGFR levels (indicative of kidney dysfunction) were associated with higher plasma concentrations of phosphorylated and unphosphorylated tau peptides measured simultaneously using the tau immunoprecipitation mass spectrometry (IP-MS) assay. (medscape.com)

If biotin is found to be a more effective labeling alternative, it could result in many improvements of safety and availability of blood products in radiological or nuclear mass casualty incidents (MCI). (uc.edu)

Isotope ration mass spectrometry was used to quantify metabolically produced nitrogen-15. (cdc.gov)

Their ionization efficiencies for mass spectrometry (MS) detection are rather poor, whereas their retention and separation on reverse phase liquid chromatography (RPLC) are often unsatisfying. (unboundmedicine.com)

Nonsupervised construction and application of mass spectral and retention time index libraries from time-of-flight gas chromatography-mass spectrometry metabolite profiles. (mpg.de)
A typical dataset contains up to tens of thousands of features (mass-to-charge and retention time (m/z-RT) pairs). (metabolon.com)

Because these differentially labeled peptides are quantitated at the MS1 level, precursor interference isn't a problem. (genomeweb.com)

By capitalizing on our stable isotope labeled RNA phosphoramidites and triphosphates innotope offers an oligonucleotide synthesis service for our customers. (innotope.at)

Isotope-labeling is a useful technique for understanding cellular metabolism. (mdpi.com)

Anatomy11

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Chemicals and Drugs85

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Disciplines and Occupations7

Anthropology, Education, Sociology and Social Phenomena6

Technology, Industry, Agriculture1

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SILAC10

Peptide7

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Carbon7

Isobaric1

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