The Secretome Analysis by High-Throughput Proteomics and Multiple Reaction Monitoring (MRM)

2014 ◽  
pp. 323-335 ◽  
Author(s):  
Vitor M. Faça ◽  
Camila S. Palma ◽  
Daniele Albuquerque ◽  
Gabriela N. S. Canchaya ◽  
Mariana L. Grassi ◽  
...  
Keyword(s):  
High Throughput ◽  
Multiple Reaction Monitoring ◽  
Reaction Monitoring ◽  
Secretome Analysis

scholarly journals High-Throughput LC/MS/MS Analysis of Ruscogenin and Neoruscogenin in Ruscus aculeatus L.

2009 ◽  
Vol 92 (4) ◽  
pp. 1055-1059 ◽  
Author(s):  
Laurian Vlase ◽  
Béla Kiss ◽  
Georgeta Balica ◽  
Mircea Tas ◽  
Gianina Crisan ◽  
...  
Keyword(s):  
Electrospray Ionization ◽  
Mass Spectrometer ◽  
High Throughput ◽  
Ion Trap ◽  
Multiple Reaction Monitoring ◽  
Reaction Monitoring ◽  
Ionization Source ◽  
Ion Trap Mass Spectrometer ◽  
Multiple Reaction Monitoring Mode ◽  
Ionization Mode

Abstract A new, sensitive LC/MS/MS method was developed for the quantification of ruscogenin and neoruscogenin in hydrolyzed extracts from Ruscus aculeatus L. (Liliaceae). The two sapogenins were separated on a Zorbax SB-C18 column under isocratic conditions. The detection was performed in the multiple reaction monitoring mode using an ion trap mass spectrometer with an electrospray ionization source operated in positive ionization mode. For the quantification of the ruscogenin and neoruscogenin, calibration curves were constructed over the range of 21000 ng/mL. This is the first reported LC/MS/MS method for the simultaneous analysis of ruscogenin and neoruscogenin, and it showed superior sensitivity when compared with other assays described in the literature. The method has been successfully applied to quantify the two sapogenins in aerial (phylloclades) and underground parts (rhizomes, roots) of Ruscus aculeatus L.

scholarly journals MitoPlex: A Targeted Multiple Reaction Monitoring Assay for Quantification of a Curated Set of Mitochondrial Proteins

2019 ◽  
Author(s):  
Aleksandr B. Stotland ◽  
Weston Spivia ◽  
Amanda Orosco ◽  
Allen M. Andres ◽  
Roberta A. Gottlieb ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Skeletal Muscle ◽  
High Throughput ◽  
Metabolic Flux ◽  
Multiple Reaction Monitoring ◽  
Mitochondrial Protein ◽  
Carbon Chain ◽  
Reaction Monitoring ◽  
In Vivo Models ◽  
Central Carbon

SummaryMitochondria are the major source of cellular energy (ATP), as well as critical mediators of widespread functions such as cellular redox balance, apoptosis, and metabolic flux. Methods to quantify mitochondrial content are limited to low throughput immunoassays, measurement of mitochondrial DNA, or relative quantification by untargeted mass spectrometry. Here, we present a high throughput, reproducible and quantitative mass spectrometry multiple reaction monitoring based assay of 37 proteins critical to central carbon chain metabolism and overall mitochondrial function termed ‘MitoPlex’. We coupled this protein multiplex with a parallel analysis of the central carbon chain metabolites (218 metabolite assay) extracted in tandem from the same sample, be it cells or tissue. In tests of its biological applicability in cells and tissues, ‘MitoPlex plus metabolites’ indicated profound effects of HMG-CoA Reductase inhibition (e.g., statin treatment) on mitochondria of i) differentiating C2C12 skeletal myoblasts, as well as a clear opposite trend of statins to promote mitochondrial protein expression and metabolism in heart and liver, while suppressing mitochondrial protein and ii) aspects of metabolism in the skeletal muscle obtained from C57Bl6 mice. Our results not only reveal new insights into the metabolic effect of statins in skeletal muscle, but present a new high throughput, reliable MS-based tool to study mitochondrial dynamics in both cell culture and in vivo models.

Mass Spectrometry in High-Throughput Clinical Biomarker Assays: Multiple Reaction Monitoring

2012 ◽  
pp. 117-137 ◽  
Author(s):  
Carol E. Parker ◽  
Dominik Domanski ◽  
Andrew J. Percy ◽  
Andrew G. Chambers ◽  
Alexander G. Camenzind ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Multiple Reaction Monitoring ◽  
Reaction Monitoring ◽  
Clinical Biomarker
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