The Experts below are selected from a list of 321 Experts worldwide ranked by ideXlab platform
Peter Sporns - One of the best experts on this subject based on the ideXlab platform.
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immunoaffinity sample Purification and maldi tof ms analysis of α solanine and α chaconine in serum
Journal of Agricultural and Food Chemistry, 2001Co-Authors: Darcy R Driedger, Peter SpornsAbstract:A sample Purification Technique was developed for the detection of potato glycoalkaloids (GAs) in blood serum by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-...
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immunoaffinity sample Purification and maldi tof ms analysis of α solanine and α chaconine in serum
Journal of Agricultural and Food Chemistry, 2001Co-Authors: Darcy R Driedger, Peter SpornsAbstract:A sample Purification Technique was developed for the detection of potato glycoalkaloids (GAs) in blood serum by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). GAs were extracted from spiked serum (5 mL) using a C18 solid-phase extraction cartridge. The GAs were then selectively captured on antibody-coated agarose beads. The agarose beads were washed with water and the GAs eluted with 25 μL of methanol. MALDI-TOF MS was used to detect the GAs in the methanol eluent. Immunoaffinity sample Purification of the GAs effectively reduced the signal suppression observed during the analysis of unpurified samples. α-Chaconine and α-solanine were detected in serum spiked with 1 ng/mL of each GA. Keywords: Potato; glycoalkaloid; solanidine; antibody; affinity column; solid-phase extraction
Darcy R Driedger - One of the best experts on this subject based on the ideXlab platform.
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immunoaffinity sample Purification and maldi tof ms analysis of α solanine and α chaconine in serum
Journal of Agricultural and Food Chemistry, 2001Co-Authors: Darcy R Driedger, Peter SpornsAbstract:A sample Purification Technique was developed for the detection of potato glycoalkaloids (GAs) in blood serum by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-...
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immunoaffinity sample Purification and maldi tof ms analysis of α solanine and α chaconine in serum
Journal of Agricultural and Food Chemistry, 2001Co-Authors: Darcy R Driedger, Peter SpornsAbstract:A sample Purification Technique was developed for the detection of potato glycoalkaloids (GAs) in blood serum by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). GAs were extracted from spiked serum (5 mL) using a C18 solid-phase extraction cartridge. The GAs were then selectively captured on antibody-coated agarose beads. The agarose beads were washed with water and the GAs eluted with 25 μL of methanol. MALDI-TOF MS was used to detect the GAs in the methanol eluent. Immunoaffinity sample Purification of the GAs effectively reduced the signal suppression observed during the analysis of unpurified samples. α-Chaconine and α-solanine were detected in serum spiked with 1 ng/mL of each GA. Keywords: Potato; glycoalkaloid; solanidine; antibody; affinity column; solid-phase extraction
Angeliki Malliri - One of the best experts on this subject based on the ideXlab platform.
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a modified tandem affinity Purification Technique identifies that 14 3 3 proteins interact with tiam1 an interaction which controls tiam1 stability
Journal of Proteome Research, 2009Co-Authors: Simon A Woodcock, Richard C Jones, Ricky D Edmondson, Angeliki MalliriAbstract:The Rac-specific GEF (guanine-nucleotide exchange factor) Tiam1 has important functions in multiple cellular processes including proliferation, apoptosis and adherens junction maintenance. Here we describe a modified tandem affinity Purification (TAP) Technique that we have applied to specifically enrich Tiam1-containing protein complexes from mammalian cells. Using this Technique in conjunction with LC-MS/MS mass spectrometry, we have identified additional Tiam1-interacting proteins not seen with the standard Technique, and have identified multiple 14-3-3 family members as Tiam1 interactors. We confirm the Tiam1/14-3-3 protein interaction by GST-pulldown and coimmunoprecipitation experiments, show that it is phosphorylation-dependent, and that they colocalize in cells. The interaction is largely dependent on the N-terminal region of Tiam1; within this region, there are four putative phospho-serine-containing 14-3-3 binding motifs, and we confirm that two of them (Ser172 and Ser231) are phosphorylated in cells using mass spectrometry. Moreover, we show that phosphorylation at three of these motifs (containing Ser60, Ser172 and Ser231) is required for the binding of 14-3-3 proteins to this region of Tiam1. We show that phosphorylation of these sites does not affect Tiam1 activity; significantly however, we demonstrate that phosphorylation of the Ser60-containing motif is required for the degradation of Tiam1. Thus, we have established and proven methodology that allows the identification of additional protein-protein interactions in mammalian cells, resulting in the discovery of a novel mechanism of regulating Tiam1 stability.
Jeffrey E Gerst - One of the best experts on this subject based on the ideXlab platform.
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rapid an aptamer based mrna affinity Purification Technique for the identification of rna and protein factors present in ribonucleoprotein complexes
Methods of Molecular Biology, 2011Co-Authors: Boris Slobodin, Jeffrey E GerstAbstract:RNA metabolism involves regulatory processes, such as transcription, splicing, nuclear export, transport and localization, association with sites of RNA modification, silencing and decay, and necessitates a wide variety of diverse RNA-interacting proteins. These interactions can be direct via RNA-binding proteins (RBPs) or indirect via other proteins and RNAs that form ribonucleoprotein complexes that together control RNA fate. While pull-down methods for the isolation of known RBPs are commonly used, strategies have also been described for the direct isolation of messenger RNAs (mRNAs) and their associated factors. The latter Techniques allow for the identification of interacting proteins and RNAs, but most suffer from problems of low sensitivity and high background. Here we describe a simple and highly effective method for RNA Purification and identification (RaPID) that allows for the isolation of specific mRNAs of interest from yeast and mammalian cells, and subsequent analysis of the associated proteins and RNAs using mass spectrometry and reverse transcription-PCR, respectively. This method employs the MS2 coat RBP fused to both GFP and streptavidin-binding protein to precipitate MS2 aptamer-tagged RNAs using immobilized streptavidin.
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a novel mrna affinity Purification Technique for the identification of interacting proteins and transcripts in ribonucleoprotein complexes
RNA, 2010Co-Authors: Boris Slobodin, Jeffrey E GerstAbstract:Intracellular mRNA targeting and localized translation are potential determinants for protein localization. To facilitate targeting, mRNAs possess specific cis-acting sequence motifs that are recognized by trans-acting RNA-binding proteins (RBPs). While many mRNAs are trafficked, our knowledge of the RBPs involved and presence of additional transcripts within these ribonucleoprotein (RNP) complexes is limited. To facilitate the identification of RBPs and transcripts that bind to specific mRNAs, we developed RNA-binding protein Purification and identification (RaPID), a novel Technique that allows for the affinity Purification of MS2 aptamer-tagged mRNAs and subsequent detection of bound RBPs and transcripts using mass-spectometry and RT-PCR, respectively. RaPID effectively isolated specific mRNAs from both yeast and mammalian cells, and identified known mRNA-RBP interactions (e.g., ASH1-She2; β-Actin-IMP1). By isolating tagged OXA1 mRNA using RaPID, we could identify a yeast COPI subunit (i.e., Sec27) as a candidate interacting protein. This finding was strengthened by the observation that a portion of OXA1 mRNA was delocalized in a sec27-1 temperature-sensitive mutant at restrictive temperatures. Finally, RaPID could also be used to show biochemically the coexistence of different RNA species within the same RNP complex (e.g., coprecipitation of the yeast SRO7, WSC2, SEC3, and IST2 mRNAs with ASH1 mRNA) for the first time.
Mika Sillanpaa - One of the best experts on this subject based on the ideXlab platform.
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water Purification using magnetic assistance a review
Journal of Hazardous Materials, 2010Co-Authors: Ritu D. Ambashta, Mika SillanpaaAbstract:Water is a major source for survival on this planet. Its conservation is therefore a priority. With the increase in demand, the supply needs to meet specific standards. Several Purification Techniques have been adopted to meet the standards. Magnetic separation is one Purification Technique that has been adapted from ore mining industries to anti-scale treatment of pipe lines to seeding magnetic flocculent. No reviews have come up in recent years on the water Purification Technique using magnetic assistance. The present article brings out a series of information on this water Purification Technique and explains different aspects of magnetism and magnetic materials for water Purification.
