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Original title:
Protein fractionation and relative quantitation using PF 2D and ITRAQ for biomarker quest
Authors: Gadher, S. J. ; Skalníková, Helena ; Halada, Petr ; Řehulka, Pavel ; Chmelík, Josef ; Kovářová, Hana
Document type: Papers
Conference/Event: 1st Central and Eastern European proteomic Conference and 3rd Czech Proteomic Conference, Praha (CZ), 2007-10-29 / 2007-10-31
Year: 2007
Language: eng
Abstract: ProteomeLab PF 2D System - Protein Fractionation in 2 Dimensions (Beckman Coulter, Fullerton, CA, USA) has been developed to fractionate complex protein mixtures by chromatofocusing in the first dimension followed by high-resolution non-porous silica reversed phase chromatography (RP LC) in the second dimension. Despite the high-resolution power of ProteomeLab PF 2D, UV-based quantitation could be compromised due to possible co-elution of several proteins into one fraction. Hence, we present an optimized protocol for application of isobaric tags for relative and absolute quantitation (iTRAQ) and MALDI-TOF/TOF mass spectrometry to obtain quantitative data from peptides derived by tryptic digestions of intact proteins fractionated by ProteomeLab PF 2D technique. To demonstrate the feasibility of such an approach, protein expression patterns obtained from the ProteomeLab PF 2D fractionation of human T-lymphoblastic leukemia CEM cell line were utilised.
Keywords: protein fractionation; proteomics
Project no.: CEZ:AV0Z50450515 (CEP), CEZ:AV0Z40310501 (CEP), CEZ:AV0Z50200510 (CEP), GA301/05/0418 (CEP), LC07017 (CEP)
Funding provider: GA ČR, GA MŠk
Host item entry: 1st Central and Eastern European Proteomic Conference and 3rd Czech Proteomic Conference
Institution: Institute of Animal Physiology and Genetics AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0175379
Permalink: http://www.nusl.cz/ntk/nusl-40462
The record appears in these collections:
Research > Institutes ASCR > Institute of Animal Physiology and Genetics
Conference materials > Papers
Authors: Gadher, S. J. ; Skalníková, Helena ; Halada, Petr ; Řehulka, Pavel ; Chmelík, Josef ; Kovářová, Hana
Document type: Papers
Conference/Event: 1st Central and Eastern European proteomic Conference and 3rd Czech Proteomic Conference, Praha (CZ), 2007-10-29 / 2007-10-31
Year: 2007
Language: eng
Abstract: ProteomeLab PF 2D System - Protein Fractionation in 2 Dimensions (Beckman Coulter, Fullerton, CA, USA) has been developed to fractionate complex protein mixtures by chromatofocusing in the first dimension followed by high-resolution non-porous silica reversed phase chromatography (RP LC) in the second dimension. Despite the high-resolution power of ProteomeLab PF 2D, UV-based quantitation could be compromised due to possible co-elution of several proteins into one fraction. Hence, we present an optimized protocol for application of isobaric tags for relative and absolute quantitation (iTRAQ) and MALDI-TOF/TOF mass spectrometry to obtain quantitative data from peptides derived by tryptic digestions of intact proteins fractionated by ProteomeLab PF 2D technique. To demonstrate the feasibility of such an approach, protein expression patterns obtained from the ProteomeLab PF 2D fractionation of human T-lymphoblastic leukemia CEM cell line were utilised.
Keywords: protein fractionation; proteomics
Project no.: CEZ:AV0Z50450515 (CEP), CEZ:AV0Z40310501 (CEP), CEZ:AV0Z50200510 (CEP), GA301/05/0418 (CEP), LC07017 (CEP)
Funding provider: GA ČR, GA MŠk
Host item entry: 1st Central and Eastern European Proteomic Conference and 3rd Czech Proteomic Conference
Institution: Institute of Animal Physiology and Genetics AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0175379
Permalink: http://www.nusl.cz/ntk/nusl-40462
The record appears in these collections:
Research > Institutes ASCR > Institute of Animal Physiology and Genetics
Conference materials > Papers
Record created 2011-07-01, last modified 2024-01-26