PMID: 11921441Mar 29, 2002Paper

Functional proteomics using microchannel plate detectors

Proteomics
Paul Richards, J E Lees

Abstract

We describe the development of a novel detection system used for the functional imaging of proteins separated on electrophoretic gels. A microchannel plate detector is used here for real-time imaging of low levels of tritiated protein separated by two-dimensional (2-D) electrophoresis. The system employs radioisotope-free, low noise microchannel plates originally developed for photon counting in X-ray astronomy. Using the detector configuration described here, proteins were resolved on mini gels by either one or two-dimensional electrophoresis, transferred onto polyvinylidene difluoride membranes and directly imaged. Tritiated diisopropylfluorophosphate (DFP) was used as a selective label for the serine hydrolase class of enzymes and their distribution in the central nervous system was examined. This survey revealed approximately 24 protein spots by 2-D electrophoresis. We also investigated the relative sensitivity of these proteins towards DFP and found the peptidase, acylpeptide hydrolase to be the most sensitive brain protein towards this reagent. Using a number of different tritiated standards, it was found that the system can image as little as 0.1 Bq/mm(2) of tritium corresponding to 320 attomol of DFP labelled protein/mm...Continue Reading

Citations

Aug 28, 2003·Current Opinion in Chemical Biology·Jonathan S Rosenblum, John W Kozarich
Mar 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Helena WrightVilmos Fülöp
Dec 7, 2010·Journal of Pharmaceutical and Biomedical Analysis·Dirk Steinmann, Markus Ganzera
Nov 3, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Wayne G CarterDavid E Ray
Jul 21, 2015·Applied Optics·Liisa M HirvonenKlaus Suhling
Apr 29, 2006·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·Satoshi Nishizuka
Jul 7, 2007·Human & Experimental Toxicology·Wayne G CarterDavid E Ray
Aug 23, 2006·The Journal of Biological Chemistry·Vasanthy VigneswaraWayne G Carter
May 20, 2008·Biomarkers : Biochemical Indicators of Exposure, Response, and Susceptibility to Chemicals·Mabruka H TarhoniWayne G Carter

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