University of the basque country UPV/EHUScience and technology faculty Edificio CD4Maria Goyri research building, CBA at UPV/EHU science parkBº Sarriena, s/n 48940 Leioa (Vizcaya)
Dr. Kerman Aloria Phone: 94 601 3278 e-mail: email@example.com
Access to the unit of Proteomics involves meeting the requirements set forth in the Protocol for access to SGIker and the services it provides.
Proteomics studies the protein - the set of proteins that is expressed via a genome. The study of the genome has produced a great deal of necessary information, albeit insufficient for knowledge about biological processes, and has made it necessary to specifically study the protein. The proteome is a highly-dynamic element that varies according to the different states and conditions in which a cell or subcellular compartment can be found. The same genome gives rise to the expression of different proteins, depending on the cellular state, while these in turn may be modified differently in response to different requirements. This means that different proteomes can be obtained from the same genome. Huge advances have been made in recent times in the study of the proteome thanks to the complete sequencing of a greater number of genomes, among them the human one; the development of powerful bioinformatic tools that enable a large amount of information to be analysed in a short time, and the technological advances - especially in mass spectrometry - which have considerably increased the quantity and quality of information obtained in experimental analysis. The Proteomics Unit of the La UPV/EHU, located at the Faculty of Science and Technology on the Bizkaia Campus, is a general service that lends support to research and whose aim is to offer its services to research groups from the UPV/EHU and to the scientific community in general. The Proteomics Unit of the UPV/EHU attempts to provide full support both on an experimental and intellectual level to its users, for which purpose we have state-of-the-art equipment and qualified personnel at our disposal.
The Proteomics Unit of the UPV/EHU offers the following services: Molecular mass analysis: the molecular mass analysis of proteins via mass spectrometry has become a routine methodology for analysing possible alterations in purified proteins whose molecular mass is known (i.e. overexpression). 2 types of analysis are carried out, depending on the complexity of the sample: Purified protein: where necessary, the buffer of the sample is cleaned and changed using Micro Bio-Spin 6 columns (Bio-Rad) Protein mixture: proteins from the sample are separated in a 5-micra Symmetry 300 C4 3.5 column (Waters). Protein identification: tandem mass spectrometry offers the chance to identify the proteins in samples of very variable complexity, ranging from purified proteins (chromatographic purifications, bidimensional gel spots) to samples of variable complexity (majority bands of SDS-PAGE gels, relatively purified chromatographic fractions, immunoprecipitations, subcellular organelles and total extracts of cells). Different types of analysis are offered, depending on the complexity of the sample and the results required: Samples with less than 5 proteins PMF - Peptide Mass Fingerprinting: MS (MALDI-Q-TOF) PMF - Peptide Mass Fingerprinting: MS + MS/MS (MALDI-Q-TOF) nanoLC-MS/MS (ESI-Q-TOF) Samples with less than 50 proteins Samples with more than 50 proteins (the sample is analysed in triplicate) Differential proteomics: the protein expression is compared in different samples via differential protein expression analysis (healthy vs. sick), differentially-expressed proteins are identified, and their variation is determined. The method used is based on liquid chromatography analysis together with mass spectrometry and without sample marking. This technique enables an unlimited number of samples to be compared. 3 replicas of each sample are required. Characterization of post-translational modifications: phosphorylation analysis is offered, in which the phosphopeptides of the sample are enriched via chromatography with TiO2 and analysis is carried out using mass spectrometry. The phosphopeptide can be identified and the amino acid phosphorylate determined. 3 types of analysis are offered, depending on the complexity of the sample: Samples with less than 5 proteins Samples with less than 50 proteins Samples with less than 50 samples Image analysis and differential quantification of 2-D gels: the Proteomics Unit of the UPV/EHU makes Progenesis SameSpot image analysis software available to users (Nonlinear Dynamics). Self-service: following a small introduction, the user then carries out the entire analysis of their images. The user provides the images to be analysed and the Proteomics Unit carries out the entire analysis. Once the sample has been analysed, the Proteomics Unit of the UPV/EHU will draft a complete report of the results obtained or any additional explanation that the user may need.