Quantification stable isotope labeled protein. Medical search. Frequent questions

Our method allows for the identification of the proteins, identification of redox-sensitive cysteines within proteins, and quantification of the redox status of individual cysteine-containing peptides. (hindawi.com)
Here we discuss two approaches currently used to investigate the global dynamics of phosphorylation based on the isolation of phosphorylated proteins or peptides. (acs.org)
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)
Absolute quantification of protein and post-translational modification abundance with stable isotope-labeled synthetic peptides. (kettenbachlab.org)
Stable isotope labeled peptides, or SIL peptides, maintain identical amino acid sequence with the natural ones released from the parent proteins by chemical or enzymatic cleavage, but some of the amino acid residuals are labeled by specific stable isotope, such as 2 H, 13 C, and/or 15 N atom(s). (novoprolabs.com)
Stable isotope labeled peptides have been widely applied in the nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS). The combination of SIL peptides with NMR spectroscopy allow for the incorporation of NMR active nuclei which can reduce the complexity of spectra, and then help researchers to obtain novel correlations between atoms for more structural information. (novoprolabs.com)
Meanwhile, the utilization of SIL peptides in quantitative proteomics can facilitate the comparison of different proteomes, and protein-protein interaction studies as internal standards, even MRM/SRM-based absolute quantification of specific target proteins in biological samples. (novoprolabs.com)
With more than 10 years of experience, and as a world-leading peptide manufacturing organization, NovoPro can provide customized peptides labeled with stable isotopes including 2 H, 15 N & 13 C, respectively, or a combination of 15 N & 13 C. Even when the peptides are attached with specific modification motifs. (novoprolabs.com)
Below lists is the most common stable isotopically labeled amino acids that can be used for the synthesis of peptides with stable isotopes containing carbon 13 ( 13 C), Nitrogen 15 ( 15 N). (novoprolabs.com)
Generally, selective quantification of antibody drugs is done using unique peptides from variable regions (V H and V L ) as surrogate peptides. (elsevierpure.com)
Further, numerous internal standards (ISs) such as stable isotope-labeled (SIL)-intact proteins and SIL-surrogate peptides are used. (elsevierpure.com)
Labels such as isobaric chemical tags with different masses can be introduced through covalent modification to peptides. (openaccesspub.org)
Multiplexed proteomics is a powerful tool to assay cell states in health and disease, but accurate quantification of relative protein changes is impaired by interference from co-isolated peptides. (biorxiv.org)
Because the overall number of heavy isotopes is kept constant between the different tags, they add the same total mass to the peptides, and hence are isobaric. (biorxiv.org)
This information is used for determining which condition the ions stem from and permits relative quantification of peptides. (biorxiv.org)
If MS2 reporter ions are used for quantification, the interfering peptides lead to a distortion of the measured ratios, as the source of reporter ions (red squares) cannot be distinguished. (biorxiv.org)
Excellent R&D teams, advanced instruments and rich production experience enable us to provide high-quality products and custom synthesis of peptides/proteins. (creative-proteomics.com)
Or you can also inquire about our comprehensive custom synthesis of peptides and proteins. (creative-proteomics.com)
The PrESTs are then spiked into cell lysates and the SILAC ratios of PrEST peptides to peptides from endogenous target proteins yield their cellular quantities. (scilifelab.se)
The workflow relies on stable isotope labelled reference peptides and selected reaction monitoring mass spectrometry analysis of a wild-type strain and an M-protein deficient mutant strain to generate absolutely quantified protein stoichiometry ratios between S. pyogenes and interacting plasma proteins. (malmstroem.net)
The quantitative analysis of proteins/peptides is not a routine service. (mslab-ibb.pl)

The issue of abundant (often housekeeping) proteins masking regulatory proteins of lower abundance (such as signaling proteins and cytokines) continues to be a challenging issue for proteomics particularly in the case of biofluids. (hindawi.com)
While modern proteomics experiments permit the analysis of hundreds to thousands of proteins in complex samples, the most powerful use of this data would combine information on protein activity with the identities of the active proteins. (hindawi.com)
Redox proteomics has taken advantage of the thiol specificity of ICAT reagents not only to identify targets of ROS but also to quantify oxidative thiol modifications in individual proteins. (hindawi.com)
Absolute Quantification (Protein-AQUA™™) is a targeted quantitative proteomics technique that exhibits robust efficacy and is being increasingly utilized for a wide variety of quantitative proteomics studies. (sigmaaldrich.com)
Discovery Proteomics and Absolute Protein Quantification Can Be Performed Simultaneously on an Orbitrap-Based Mass Spectrometer. (ncsu.edu)
Therefore, the development of methods for accurate protein quantitation is one of the most challenging areas of proteomics. (openaccesspub.org)
Cristian Ruse, Emily I. Chen (2014) Discovery and Quantification in Mass Spectrometry-Based Proteomics. (openaccesspub.org)
Scientists from Creative Proteomics offer you the high-density microarrays with thousands of unique full-length proteins that can be detected at the same time. (creative-proteomics.com)
Creative Proteomics is a leading proteomics company offering SIL protein/peptide standards, protein microarrays and other related products for highly multiplexed protein quantification techniques. (creative-proteomics.com)
ISOTEC® Stable Isotopes are stable isotope compounds, ranging from gases to complex molecules, that are useful for tracer studies in proteomics and metabolomics, agents for MRI / MRS, and in a wide range of other biomedical applications. (sigmaaldrich.com)
To confirm differential proteins that are revealed by the above techniques and develop protein diagnostic panels, targeted proteomics methods based on multiple reaction monitoring/parallel reaction monitoring (MRM/PRM) data acquisition protocols are applied. (edu.pl)
Mass spectrometry-based proteomics increasingly relies on relative or absolute quantification. (scilifelab.se)
Targeted proteomics and absolute protein quantification for the construction of a stoichiometric host-pathogen surface density model. (malmstroem.net)
Here, we employ stable isotope labeling with amino acids in cell culture (SILAC)-based proteomics to quantify the response of over 6200 proteins to increasing concentrations of tunicamycin in HeLa cells. (torvergata.it)

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)
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)
A Protein Epitope Signature Tag (PrEST) library allows SILAC-based absolute quantification and multiplexed determination of protein copy numbers in cell lines. (scilifelab.se)
We first quantify a highly purified, stable isotope labeling of amino acids in cell culture (SILAC)-labeled version of the solubility tag and use it determine the precise amount of each PrEST by its SILAC ratios. (scilifelab.se)
Direct quantification of the absolute amount of single proteins is possible via a SILAC experiment in which labeled cell lysate is mixed both with the heavy labeled solubility tag and with the corresponding PrEST. (scilifelab.se)
The SILAC-PrEST combination allows accurate and streamlined quantification of the absolute or relative amount of proteins of interest in a wide variety of applications. (scilifelab.se)
For the training dataset, HEK cells were grown in either light or heavy SILAC (Stable Isotope Labeling with amino acids in cell culture) medium, lysed and mixed together in a certain ratio. (sched.com)
Methods requiring stable isotope labeling of samples (iTRAQ, SILAC), as well as label-free approaches, can be used for this type of quantification and are selected individually in accordance with experiment design. (mslab-ibb.pl)

The small ubiquitin-like modifiers, SUMO-1, SUMO-2 and SUMO-3 are conjugated to a wide, but poorly defined range of target proteins and play important roles in many cellular processes. (europa.eu)
Employment of most modern proteomic and bioinformatic techniques and tools allowed us to undertake the large-scale identification of SUMO-1 and SUMO-2 putative target proteins. (europa.eu)
Extensive analysis of obtained results resulted in much deeper understanding of how poly-SUMO-2 chains are being assembled into longer forms, with preference for extending of already existing chains on target proteins rather than recruitment of large amounts of new substrates. (europa.eu)
To identify CAMK2D target proteins in renal collecting duct cells (mpkCCD), we deleted Camk2d and carried out LC-MS/MS-based quantitative phoshoproteomics. (bvsalud.org)
This adds to the list of known CAMK2D target proteins found in prior reductionist studies. (bvsalud.org)
Multiple primary PCSs were used for quantification of many target proteins. (elsevierpure.com)
Furthermore, we varied the copy number of the "ID-tag peptide" in each PCS according to a range of expression levels of target proteins. (elsevierpure.com)

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)
This methodology enables detection of putative metabolites from biological samples and simultaneous quantification of the pattern and extent of isotope labeling. (acs.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)
We routinely design and execute multi-step syntheses of complex labeled materials, including metabolites and steroids, fatty acids, amino acids, reference standards, pharmaceutical intermediates and other bioactive compounds. (sigmaaldrich.com)
The metabolomic division of our facility allows small-molecule quantification, with a special focus on drugs and their metabolites. (edu.pl)
Quantification of Stable Isotope Label in Metabolites via Mass Spectrometry. (mpg.de)

However, because of the unknown binding affinity of antibody to MRP2/Mrp2 protein across species and lack of purified MRP2/Mrp2 proteins for calibration, the immunoblotting assay was excluded from the absolute quantification of MRP2/Mrp2 protein for multiple species. (aspetjournals.org)
Understanding K. phaffii (Pichia pastoris) Host Cell Proteins: Proteomic Analysis and Flow-through Affinity Clearance (T. I. Williams, Ed.). Biotechnology and Bioengineering, Vol. 01. (ncsu.edu)
Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via "flow-through affinity chromatography" using peptide-based adsorbents. (ncsu.edu)
Unlike immunofluorescence microscopy approaches, which provide correlative information about proteins sharing cellular space, co-IP can confirm direct often high-affinity physical interactions between two proteins or protein complexes of equal or better affinity. (aacrjournals.org)

The variability introduced by different methods prior to quantitative mass spectrometry should be considered, and accuracy and precision of MS measurements can also vary depending on the strategy used for MS quantification. (openaccesspub.org)

Here we presented a strategy by integration of the stable isotope dimethyl labeling with variation-associated database search to globally quantify the SAAVs at the first time. (nih.gov)

Improvements in mass spectrometry has led to the development of a number of techniques to quantify the relative protein abundance within a given sample. (hindawi.com)
AQP2 protein abundance was lower in the CAMK2D knockout cells than in CAMK2D-intact controls. (bvsalud.org)
Large-scale quantitative phosphoproteomic analysis (TMT-labeling) in the presence of the vasopressin analog dDAVP (0.1 nM, 30 min) allowed quantification of 11,570 phosphosites of which 169 were significantly decreased, while 206 were increased in abundance in CAMK2D knockout clones. (bvsalud.org)
The accurate and precise absolute abundance of proteins can be determined using mass spectrometry by spiking the sample with stable isotope-labeled standards. (elsevierpure.com)
The quantified abundance of budding yeast proteins showed a high reproducibility for replicate analyses and similar copy numbers per cell for ribosomal proteins, demonstrating the accuracy and precision of this strategy. (elsevierpure.com)
Thus, the approach presented here is a faithful method for the absolute quantification of proteomes and provides insights into biological mechanisms, including the regulation of expressed protein abundance. (elsevierpure.com)
For each experiment, data validation and statistical analysis is performed, to obtain a list of proteins which show a significant abundance change between control and experimental samples. (mslab-ibb.pl)

03 MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. (imarkerlab.com)
Using these quantitative approaches, one can investigate the dynamics of proteome through differential protein expression in normal biological processes and diseases. (openaccesspub.org)
This type of analysis provides selective quantitative information on the proteins of interest and is less time-consuming than global analysis of a whole proteome. (mslab-ibb.pl)
The unfolded protein response (UPR) involves extensive proteome remodeling in many cellular compartments. (torvergata.it)
This systematic quantification of the proteome-wide expression changes that follow proteostatic stress is a resource for the scientific community, enabling the discovery of novel players involved in the pathophysiology of the broad range of disorders linked to proteostasis. (torvergata.it)
mHTT-induced proteome and protein interaction alterations have been investigated in the brain, yet those proximal to disease progression remain poorly understood. (skyline.ms)

Here we used the gram-positive bacterium Streptococcus pyogenes as a model system to establish a mass spectrometry based workflow for the construction of a stoichiometric surface density model between the S. pyogenes surface the surface virulence factor M-protein and adhered human blood plasma proteins. (malmstroem.net)
We present a fully integrated workflow for peptide identification and quantification within Skyline that incorporates the MS Amanda search algorithm. (skyline.ms)

We describe the design and synthesis of a novel set of iso-baric tags for quantitative glycan profiling, which will have broad applications in carbohydrate based biomarker dis-covery, therapeutic protein characterization, and vaccine development. (nist.gov)
Our ISOTEC ® Stable Isotopes team specializes in the custom synthesis of labeled compounds, including the manufacturing, testing, and control of cGMP (current good manufacturing practices) materials. (sigmaaldrich.com)

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)
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)
Here, we report the use of a targeted liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) method for concurrent quantification of multiple cytochrome P450 (P450), UDP-glucuronosyltransferase (UGT), and transporter proteins in cultured primary human hepatocytes. (aspetjournals.org)
10 A statistical model for identifying proteins by tandem mass spectrometry. (imarkerlab.com)
This method required 10 microliters of serum and utilizes liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and isotope dilution for quantification. (cdc.gov)
Fazili Zand Pfeiffer CM. Accounting for an isobaric interference allows correct determination of folate vitamers in serum by isotope dilution-liquid chromatography-tandem mass spectrometry external icon . (cdc.gov)
Fazili Z, Pfeiffer CM. Measurement of folates in serum and conventionally prepared whole blood lysates: application of an automated 96-well plate isotope-dilution tandem MS method external icon . (cdc.gov)
McCoy LF, Scholl PF, Schleicher RL, Groopman JD, Powers CD, Pfeiffer CM. Analysis of aflatoxin B1-lysine adduct in serum using isotope-dilution liquid chromatography/tandem mass spectrometry external icon . (cdc.gov)
Mineva EM, Schleicher RL, Chaudhary-Webb, M, May KL, Botelho JC, Vesper HW, Pfeiffer CM. A candidate reference measurement procedure for quantifying serum concentrations of 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 using isotope-dilution liquid chromatography-tandem mass spectrometry external icon . (cdc.gov)

Whereas changes in the levels of reduced cysteine between samples undergoing a short-term treatment can be compared immediately (Fig. 1a ), different cell lines or tissues derived from different mice have distinct proteomes and require normalisation for protein levels. (filekav.com)
05 In-gel digestion for mass spectrometric characterization of proteins and proteomes. (imarkerlab.com)

A protein database containing 87,745 amino acid variant sequences and 73,910 UniProtKB/Swiss-Prot canonical protein entries was constructed for database search, and higher energy collisional dissociation combined with collision-induced dissociation fragmentation modes were applied to improve the quantification coverage of SAAVs. (nih.gov)
We find that the beneficial fragmentation properties of TMTPro increase quantification signal five-fold compared to TMT. (biorxiv.org)

The first applications of this technology involved exposing purified proteins to either OS or normal condition before labeling with either heavy or light ICAT reagents, respectively. (hindawi.com)
09 Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents. (imarkerlab.com)
2 , 4 The different quantification channels are encoded by the distribution of heavy isotopes between the reporter and balancer region of the reagents. (biorxiv.org)

Targeted capture of Chinese hamster ovary host cell proteins: Peptide ligand binding by proteomic analysis. (ncsu.edu)
G lobal relative proteomic analysis - is performed for complex protein mixtures (body fluids, cell compartments etc.) on the Orbitrap Velos mass spectrometers. (mslab-ibb.pl)

To demonstrate the power of TMTProC, we analyzed a human and yeast interference sample and were able to quantify 13,290 proteins in 24 fractions. (biorxiv.org)
In this study, we developed a strategy of hierarchical use of peptide-concatenated standards (PCSs) to quantify more proteins over a wider dynamic range. (elsevierpure.com)

The variation rates in different samples were evaluated, and some interesting SAAVs with significant increase normalized quantification ratios, such as T1406N in CPS1 and S197R in HTATIP2, were observed to highly associate with HCC progression. (nih.gov)
As shown in Fig. 1b , a fraction of the lysates used to prepare IAM-labelled samples are digested and dimethylated with either light (H 12 CHO/NaBH 3 CN) or heavy (D 13 CDO/NaBD 3 CN) formaldehyde/sodium cyanoborohydride, mixed in equal ratios using a label-swap replication strategy for independent replicates, and subjected to high pH reversed phase chromatography fractionation before UHPLC-MS/MS analysis. (filekav.com)
This strategy accomplished absolute quantification over a wider range than that of the measured ratios. (elsevierpure.com)
The stoichiometry ratios in combination with a novel targeted mass spectrometry method to measure cell numbers enabled the construction of a stoichiometric surface density model using protein structures available from the protein data bank. (malmstroem.net)

Pawlosky RJ, Flanagan VP, Pfeiffer CM. Determination of 5-methyltetrahydrofolic acid in human serum by stable-isotope dilution high-performance liquid chromatography-mass spectrometry external icon . (cdc.gov)

Cysteine is the most important redox-responsive amino acid within proteins largely due to the wide range of oxidation states that sulfur can occupy-so called, "sulfur switches" [ 4 ]. (hindawi.com)
Global quantification of the single amino-acid variations (SAAVs) is essential to investigate the roles of SAAVs in disease progression. (nih.gov)
The separation of the 17 amino acids is performed using pre-column derivatization (OPA/FMOC derivatives) and reversed-phase chromatography (Hypersil AA-ODS) according to Agilent Application Note 5968-5658E (Angelika Gratzfeld-Huesgen, Sensitive and Reliable Amino Acid Analysis in Protein Hydrolysates using the Agilent 1100 Series HPLC). (sigmaaldrich.com)

Complex biological samples have also been successfully separated in multiple dimensions at the protein level. (hindawi.com)
Traditional measurement of the relative absolute quantity of a protein between two samples is not always necessarily proportional to the activity of the protein. (hindawi.com)
However, measuring the relative quantity of a protein between two samples does not tell us anything about the activity of the protein itself. (hindawi.com)
This method allowed for the precise quantification of targeted metabolite biomarkers using an OC blood cohort comprising control samples (n = 464), benign samples (n = 3), and OC samples (n = 13).Eleven functional modules were identified as significant differentiators (false discovery rate, FDR (stanford.edu)
By combining the results from two-stage quantification, 4 or 5 orders of magnitude in dynamic range were achieved for the detection of the serotypes of BoNT/A, /B, /E, or /F. The effect of multiplexing the assay by mixing substrates for different BoNT serotypes into a single reaction was also investigated in order to reduce the numbers of the cleavage reactions and to save valuable clinical samples. (cdc.gov)
Control and oxidatively stressed cells or tissue samples were extracted separately in the presence of either light ( 12 C 2 H 4 INO) or stable isotope-labelled heavy ( 1step three C 2 D 2 H 2 INO) IAM to alkylate reduced cysteine thiols (SH), coupling a carbamidomethyl (CAM) group to the cysteine residue. (filekav.com)
Quantitative measurements derived from the MS technology can be translated into differential expression of proteins in biological samples, and collective molecular changes can then be used to describe complex phenotypes such as disease progression. (openaccesspub.org)
b ) Volcano plot showing the distribution of proteins according to their enrichment in crosslinked (CL) over non-CL samples. (nature.com)
d ) Validation of the yeast mRNA interactome using western blotting of input samples and eluate after interactome capture with specific antibodies (ADH1, alcohol dehydrogenase 1, PUB1) or against TAP-tagged proteins (PGK1, phosphoglycerate kinase 1, TDH1, triose phosphate dehydrogenase, TRX2, thioredoxine 2, SHE2, Swi5p-dependent HO Expression 2). (nature.com)
Recently, the data-independent acquisition (DIA) analysis has been added to the spectrum of our offered methods, which enables protein quantification in complex samples in a more accurate manner than the aforementioned data-dependent techniques. (edu.pl)
Quantitative analysis performed in the Laboratory allows either to measure the absolute amount of a particular protein, or to compare the protein levels in two or more groups of biological samples. (mslab-ibb.pl)
Briefly, calibrators, quality controls, a matrix blank, blind QCs, and NHANES samples were prepared by performing a protein digestion using pronase to release the chlorinated tyrosine biomarkers. (cdc.gov)

Absolute quantification with mass spectrometry and isotope labelled internal standards has found broad applications in biomedical research. (nist.gov)

The cross-reactions of MRP2/Mrp2 protein of different species with anti-human MRP2 polyclonal antibody were found by Western blotting. (aspetjournals.org)
We took advantage of the antibody resources publicly available from the Human Protein Atlas project covering more than 80% of all human protein-coding genes. (nih.gov)

Epitope mapping revealed that a majority of the polyclonal antibodies recognized multiple linear epitopes, and based on these results, a semi-automated method was developed for peptide enrichment using polyclonal antibodies immobilized on protein A-coated magnetic beads. (nih.gov)
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)
Here, we show that we were able to use small amounts of antibodies (50 ng per target) in this manner for efficient multiplex analysis of quantitative levels of proteins in a human HeLa cell lysate. (nih.gov)

Proteomic approaches have proven powerful at identifying large numbers of proteins, but there are fewer reports of functional characterization of proteins in biological tissues. (hindawi.com)
Mass spectrometric approaches to protein structure characterization are offered, including hydrogen-deuterium exchange, X-linking, native mass spectrometry, and ion mobility. (edu.pl)
Proteomic services include protein mass determination, protein identification from gel or solution, qualitative and semi-quantitative characteristics of the protein interactome, and the characterization and mapping of a wide spectrum of posttranslational modifications. (edu.pl)

These proteins may modulate the interaction of platelets with their local cellular environment. (hindawi.com)
Ca2+/Calmodulin-dependent protein kinase 2 (CAMK2) family proteins are involved in regulation of cellular processes in a variety of tissues including brain, heart, liver and kidney. (bvsalud.org)
In other scenarios, quantitative measurements derived from the MS technology can be used to associate cooperative protein expression changes with cellular responses such as cell signaling. (openaccesspub.org)
RNA-binding proteins (RBPs) mediate pivotal cellular functions such as RNA transport, degradation or translation and represent key effectors of post-transcriptional gene regulation. (nature.com)
Robinson, Margaret S dc.description.abstract: Vesicluar transport of proteins from endosomes to the trans-Golgi network (TGN) is an essential cellular pathway, but much of its machinery is still unknown. (cam.ac.uk)
This involves purification of a specific protein from the pool of cellular proteins, followed by Western blotting of a suspected interacting protein ( 1 ). (aacrjournals.org)

Improving therapeutic protein secretion in the probiotic yeast Saccharomyces boulardii using a multifactorial engineering approach. (ncsu.edu)
Only 60% of yeast and 73% of the human RBPs have functions assigned to RNA biology or structural motifs known to convey RNA binding, and many intensively studied proteins surprisingly emerge as RBPs (termed 'enigmRBPs'), including almost all glycolytic enzymes, pointing to emerging connections between gene regulation and metabolism. (nature.com)
c ) Overlap of mRNA interactome proteins in yeast and HuH-7. (nature.com)
Quantitative Measurement of Allele-Specific Protein Expression in a Diploid Yeast Hybrid by LC-MS . Z. Khan , J. Bloom, S. Amini, M. Singh, D. Perlman, A. Caudy, L. Kruglyak. (princeton.edu)
REIL proteins of Arabidopsis thaliana interact in yeast-2-hybrid assays with homologs of the yeast RLP24, RPl24a, RLP24b, ARX1, and jjj1 proteins. (mpg.de)

Chaudhary-Webb M, Schleicher RL, Erhardt JG, Pendergrast EC, Pfeiffer CM. An HPLC ultraviolet method using low sample volume and protein precipitation for the measurement of retinol in human serum suitable for laboratories in low- and middle-income countries external icon . (cdc.gov)

We show that this conserved RNA interactome harbours many proteins without previously assigned roles in RNA biology (enigmRBPs), including surprisingly many metabolic enzymes. (nature.com)
As science evolves, new labeled substrates are needed in a wide range of applications, including clinical study tracers, labeled amino acids for protein quantification, and metabolic research standards. (sigmaaldrich.com)

We previously reported that hepatobiliary transporter multidrug resistance-associated protein (MRP2/Mrp2) is considered to be the major cause of the interspecies differences detected by efflux of fluorescent substrates in isolated hepatocytes. (aspetjournals.org)
Each serotype of BoNT hydrolyzes their protein substrates at different peptide bonds. (cdc.gov)

In 2002, the Nobel Prize in Chemistry was awarded to John Bennett Fenn for the development of electrospray ionization (ESI) and Koichi Tanaka for the development of soft laser desorption (SLD) and their application to the ionization of biological macromolecules, especially proteins. (wikipedia.org)
11 Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry. (imarkerlab.com)

Bioanalysis for large molecules, like Proteins or specific Biotherapeutics, refer to a set of methods, assays, and procedures that enable scientists to analyze specific proteins found in living organisms and the biochemical reactions underlying life processes. (sigmaaldrich.com)
Esmaeili R, Zhang M, Sternberg MR, Mapango C, Pfeiffer CM . A new multiplex immunoassay for serum ferritin, soluble transferrin receptor, C-reactive protein, α-1-glycoprotein, and retinol-binding protein showed variable comparability with traditional reference assays external icon . (cdc.gov)

A previous study from our laboratory using a MuDPIT (multidimensional protein identification technology) approach identified over 300 proteins secreted by platelets upon thrombin activation [ 1 ]. (hindawi.com)
In addition, the effectiveness of antitoxin treatment may vary depending on circulating toxin levels, so that rapid identification and quantification of BoNT proteins in clinical specimens and food is desirable for optimization of treatment. (cdc.gov)
This training covers protein identification and quantification based on stable isotopes. (sched.com)
As such, approaches for rapid, unbiased identification of protein interactomes are becoming a standard tool in the researchers toolbox, rather than a technique that is only available to specialists. (aacrjournals.org)
The number of studies that have used protein interaction discovery approaches for identification of associated factors is overwhelming. (aacrjournals.org)
These tend to fall roughly into two categories: validation of putative or suspected interactions and identification of unknown associated proteins. (aacrjournals.org)

Quantitative proteomic methods fall into two general categories: label-based or label-free quantification. (openaccesspub.org)
In relative quantification, stable isotope based methods often allow mixing at early stages of sample preparation, whereas for absolute quantification this has generally required recombinant expression of full length, labeled protein standards. (scilifelab.se)
There are numerous methods for the discovery of unknown associated protein networks, with purification of the bait (the protein of interest) followed by mass spectrometry as a common theme. (aacrjournals.org)
In recent years, advances have permitted the purification of endogenous proteins and methods for scaling down starting material. (aacrjournals.org)
We do not aim to provide an exhaustive analysis of all available techniques, but instead will highlight some of the most widely used methods for protein-protein assessment and will describe some recent advances in global protein-protein analyses that are enabling non-experts to identify interactomes of their favorite factor. (aacrjournals.org)

The procedure can readily be multiplexed, as we demonstrate by simultaneously determining the copy number of 40 proteins in HeLa cells. (scilifelab.se)

Sector mass spectrometers known as calutrons were developed by Ernest O. Lawrence and used for separating the isotopes of uranium during the Manhattan Project. (wikipedia.org)
Using the cutting-edge mass spectrometry-based proteomic techniques, she has been focused on elucidating mechanisms of tissue-specific breast cancer metastasis and identifying protein targets to eradicate the growth of distal breast cancer metastasis. (openaccesspub.org)
Mass spectrometry has become the method of choice for analysing proteins, demanding reliable and state-of-the-art software. (skyline.ms)

The effects of culture format (i.e., sandwich culture versus conventional culture) and of dexamethasone (DEX) media concentrations on mRNA, protein, and activity levels were determined for three donors, and protein expression was compared with that in liver. (aspetjournals.org)
We conclude that targeted absolute LC-MS/MS quantification of drug-metabolizing enzymes and transporters can broaden the scope and significantly increase the impact of in vitro drug metabolism studies, such as induction, as an important supplement or future alternative to mRNA and activity data. (aspetjournals.org)
Proteins shown in red (FDR 0.01) represent the mRNA interactome. (nature.com)
Unexpectedly, there are few strongly downregulated proteins, despite expression of the pro-apoptotic transcription factor CHOP, suggesting that IRE1-dependent mRNA decay (RIDD) has a limited contribution to ER stress-mediated cell death in our system. (torvergata.it)

Insulin stimulates the phosphorylation of the exocyst protein Sec8 in adipocytes. (kettenbachlab.org)
The absolute quantification strategy: application to phosphorylation profiling of human separase serine 1126. (kettenbachlab.org)
07 A probability-based approach for high-throughput protein phosphorylation analysis and site localization. (imarkerlab.com)

Reversible cysteine modifications allow for potential redox regulation of proteins. (hindawi.com)
In proteins where cysteine is not within the active site, activity can be modulated by changing conformation or by influencing its regulatory role, for example, iron sulfur complexes (ISCs) in aconitase possess cysteines required for its activity [ 7 ]. (hindawi.com)
Allosterically regulated proteins that require an activator are sometimes based on a thiol exchange interaction involving cysteines, for example, pyruvate kinase uses fructose bisphosphate (FBPs) as a heterotrophic activator and it contains a cysteine in its FBP binding site [ 8 ]. (hindawi.com)
Thus, reversible cysteine modifications can influence protein activity and the relative quantification of the status of the thiol can potentially provide valuable insights into protein activity where the protein exists in a range of redox states. (hindawi.com)

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 validity of co-IP approaches can quickly deteriorate when forced expression of tagged proteins are used to investigate possible interactions, yet this approach, particularly when endogenous factors are assessed, can provide useful information about potential protein associations. (aacrjournals.org)

Quantitative NMR (qNMR) offers many advantages over other analytical techniques with regard to quantification or purity determination of organic substances. (sigmaaldrich.com)

The sandwich culture format and 0.1 μM DEX in media retained the protein expression in the hepatocytes closest to the levels found in liver. (aspetjournals.org)
However, higher in vitro expression was observed for drug transporters, especially for multidrug resistance protein 1 and breast cancer resistance protein. (aspetjournals.org)
Explore protein expression patterns, functional annotations, and molecular interactions across diverse biological systems. (imarkerlab.com)
Lamin A/C cumulative protein expression of all variants is reported by semi-quantitative Western blotting. (mdpi.com)
We identified increased expression in 38 proteins not previously linked to the UPR, of which 15 likely remediate ER stress, and the remainder may contribute to pathological outcomes. (torvergata.it)

We detected 219 differential protein candidates in mHTT liver using MS1-based LFQ, which were all targeted for validation by PRM using Skyline. (skyline.ms)

These PrESTs are expressed recombinantly in E. coli and they consist of a short and unique region of the protein of interest as well as purification and solubility tags. (scilifelab.se)

BoNTs are dichain proteins consisting of a light chain of 50 kDa and a heavy chain of 100 kDa linked by a disulfide bond 5 . (cdc.gov)
While the original TMT-tag can incorporate a maximum of five heavy isotope labels (asterisks) in its balancer group, this number is increased to nine in the TMTPro balancer region. (biorxiv.org)
To make use of the increased plexing potential, the reporter group in TMTPro utilizes an iso-butylpyrrolidine-moiety, which can incorporate nine heavy isotope labels. (biorxiv.org)
However, because the masses of complementary ions are peptide-dependent and include the heavy isotope labels of the balancer-region (blue rectangles), they can be used for interference-free, accurate MS2 quantification. (biorxiv.org)
isotope labeled heavy vs. light quantification (e.g. (princeton.edu)

2010). Tyrosine residues are oxidized by hypochlorite at the ortho positions resulting in two stable chlorine adducts, 3-chlorotyrosine (Cl-Tyr) and 3,5-dichlorotyrosine (Cl2-Tyr) that can be used as biomarkers of chlorine gas exposure (Hureiki et. (cdc.gov)

An alternative approach is quantification by complementary ions, which allows accurate and precise multiplexed quantification at the MS2 level and is compatible with the most widely distributed instruments. (biorxiv.org)

After digestion, any undigested proteins were removed by protein precipitation. (cdc.gov)

The Laboratory also offers measurements of absolute protein concentration with AQUA ( AQUA - absolute quantification of proteins ) strategy. (mslab-ibb.pl)

Our highly trained group of stable isotope scientists are dedicated to supporting all your exploration needs. (sigmaaldrich.com)

Proteins identified within the fraction with monocyte promigratory activity and not in other inactive fractions included vimentin, PEDF, and TIMP-1. (hindawi.com)

SUMO is attached only to a specific lysine residue in the target protein. (europa.eu)

These toxins inhibit the release of neurotransmitters at the neuromuscular junction by cleaving soluble NSF attachment protein receptor (SNARE) complex proteins in nerve cells, which prevents the fusion of neurotransmitter containing vesicles to cell membranes 2 . (cdc.gov)
The light chain is a zinc-metalloprotease which is responsible for the cleavage of three SNARE complex proteins including SNAP-25 (Synaptosome-associated protein of 25 kDa), synaptobrevin-2 (also termed VAMP-2) and syntaxin. (cdc.gov)
A screen for genes involved in endosome-to-TGN trafficking produced two hits, the adaptor protein-1 (AP-1 complex), which facilitates vesicle budding, and WDR11. (cam.ac.uk)
Here we demonstrate that WDR11 forms a stable complex with two other proteins, which localises to the TGN region and does not appear to be associated with AP-1, suggesting it may act downstream from budding. (cam.ac.uk)

Anatomy17

Organisms31

Chemicals and Drugs77

Analytical, Diagnostic and Therapeutic Techniques and Equipment52

Psychiatry and Psychology6

Phenomena and Processes49

Disciplines and Occupations10

Anthropology, Education, Sociology and Social Phenomena6

Technology, Industry, Agriculture8

Humanities1

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Health Care25

Geographicals3

Peptides21

Proteomics14

SILAC8

Target proteins7

Metabolites7

Affinity4

Quantitative mass spectrometry1

Dimethyl1

Abundance7

Proteome6

Workflow2

Synthesis2

Tandem9

Proteomes2

Fragmentation2

Reagents3

Proteomic analysis2

Quantify2

Ratios4

Dilution1

Amino acid3

Samples12

Internal standards1

Antibody2

Antibodies3

Characterization3

Cellular6

Yeast5

Human Serum1

Metabolic2

Substrates2

Chemistry2

Assays2

Identification6

Methods5

HeLa1

Mass3

MRNA4

Phosphorylation3

Cysteine4

Standards1

Endogenous1

Determination1

Expression5

Differential1

Purification1

Heavy5

Biomarkers1

Accurate and precise1

Digestion1

AQUA1

Scientists1

Fractions1

Residue1

Complex4