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Acrylamide-agarose copolymers: improved resolution of high molecular mass proteins in two-dimensional gel electrophoresis

Roncada, Paola and Cretich, Marina and Fortin, Riccardo and Agosti, Susanna and De Franceschi, Lucia and Greppi, Gianfranco and Turrini, Francesco Michelangelo and Carta, Franco and Turri, Stefano and Levi, Marinella and Chiari, Marcella (2005) Acrylamide-agarose copolymers: improved resolution of high molecular mass proteins in two-dimensional gel electrophoresis. Proteomics, Vol. 5 (9), p. 2331-2339. eISSN 1615-9861. Article.

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DOI: 10.1002/pmic.200401272

Abstract

A method was developed in order to analyse high molecular mass proteins by two-dimensional (2-D) electrophoresis using a copolymer of acrylamide and allyl agarose instead of Bis cross-linked polyacrylamide (PA) gels in sodium dodecyl sulphate-electrophoresis. In this work, the matrix composition was optimised to improve the resolution of proteins larger than 200 kDa. The new gel type does not entrap large proteins and protein complexes at the application site. Mechanical properties were investigated through rheological measurements, which suggested the formation of a highly entangled elastomeric soft gel. A high 2-D resolution of proteins, extracted from membranes of red blood cells, was obtained in these gels. An example of tryptic digestion, peptide extraction and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry was reported. The results demonstrate that the new gel is fully compatible with mass spectrometry protein analysis.

Item Type:Article
ID Code:3901
Status:Published
Refereed:Yes
Uncontrolled Keywords:Bidimensional electrophoresis, cross-linker, electrophoresis
Subjects:Area 07 - Scienze agrarie e veterinarie > AGR/19 Zootecnica speciale
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze zootecniche
002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
Publisher:Wiley
eISSN:1615-9861
Additional Information:Paper presented at the 6th Siena Meeting, Siena, Italy, 30 August-2 September 2004.
Deposited On:18 May 2010 14:09

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