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Nowadays the Genomics and Proteomics Service Facility of Universidad Complutense de Madrid has the following equipment
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MALDI-TOF Mass Spectrometer with a MALDI ( Matrix-assisted Laser Desorption Ionization ) source of ionization and a TOF (Time of Flight) analyzer, model Voyager-DE™ STR Biospectrometry Workstation (Applied Biosystems), with a 2 GHz Acquisition System, a CID (C ollisionally-Induced Dissociation ) cell and the ability to acquire PSD spectra |
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The MALDI-TOF mass spectrometer is specially designed for the analysis and the identification of proteins by mass fingerprinting: peptides resulting from protein digestion are analysed by MALDI-TOF.
In addition, the accurate determination of molecular weight of proteins makes possible their characterization. |
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Tandem Quadrupole-Time of Flight Mass Spectrometer with an ESI (Electrospray Ionization) source of ionisation, model Q-TOF-2 (Waters), coupled with a Capillary Liquid Chromatography system (Waters). |
| The tandem Quadrupole/Time of Flight allows the sequencing of peptides coming from the tryptic digestion of a protein. This technique allows us to obtain “de novo” sequence to identify those proteins that are not annotated in any database. |
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| When the Q-TOF mass spectrometer is coupled with a liquid capillary chromatography system it is possible to separate the peptides prior detection. Different types of reverse phase columns along with nano-electrospray ionization provide us the highest sensitivity (few femtomoles), making easier the protein identification |
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Another application is protein characterization by analysing the post-translationals modifications, such as phosphorylation.At the present time an ionic exchange column can be connected to the system. The combination of both ionic exchange and reverse-phase separation led to a new method named multidimensional chromatography. This method allows us the direct analysis of complex proteins extracts, with no previous separation by bidimensional electrophoresis |
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Applied Biosystems 4700 Proteomics Analyzer with TOF/TOF™ optics is a MALDI tandem Time-Of-Flight mass spectrometer with CID (Collisionally-Induced Dissociation) for optimal peptide fragmentation. |
Protein mass fingerprinting identification is not always successful, especially in the case of species whose proteins are little represented in the databases. In this situation, MS/MS analysis of a peptide is needed to obtain the amino acids sequence and therefore, the protein identification by different methods (Searches in sequences EST Databases, homology analysis by BLAST, and de novo sequence). |
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Amersham Bioscience Digester Multiple probe 215 liquid handler and Turbo Vap 96 Workstation |
| The digester allows the automation of protein digestion. The sample can be in solution or in polyacrylamide gels (mono/bidimensional), previously stained with Coomasie Blue or with Silver solution. The digester also allows peptide extraction for mass spectrometry analysis |
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Digestor Multiple probe 215 liquid handler y Turbo Vap 96 Workstation
de Amersham Bioscience |
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Nos facilita llevar a cabo de forma automatizada la digestión de
proteínas con tripsina, en solución o bien, a partir de geles de poliacrilamida
mono- o bidimensionales, teñidos con Azul de Coomassie o con plata
y extracción de los péptidos. Así como la concentración de los péptidos
resultantes para su posterior análisis por espectrometría de masas. |
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System of automatic measurement of biomolecular interactions, Biacore 3000 |
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