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The services offered in the proteomics section of the Central Service for Experimental Research are encompassed in the kinds of services indicated below. Service request of proteomics, LIMS.

Preparation of the sample

A suitable sample preparation is crucial to the ultimate success of the proteomic analysis. In the proteomics section basic methodologies of preparation of the sample are offered in order to obtain protein or peptide extracts in optimal conditions for the subsequent protein analysis.

Two-dimensional electrophoresis in polyacrylamide gels

The two-dimensional electrophoresis is a decisive technique occasionally used in the proteome study, since it allows the separation of a large number of proteins from a complex protein sample. The isolation of proteins is based in a double separation in gel, through which proteins are separated in an early stage depending on the isoelectric point (pl), and secondly, depending on the molecular weight (MW).

The two-dimensional gels stained for their visualisation using staining techniques comptible with mass spectrometry: Colloidal Coomassie, compatible silver or fluorescence (SyproRuby).

Analysis of molecular weight

Analysis of molecular weight of different biomolecules by MALDI-TOF or qTOF mass spectrometry can be carried out in the proteomics section of the Central Service for Experimental Research.

Identification of proteins

Mass spectrometry is an analytical technique for basic use in the field of proteomics, which allows to determine, with high accuracy and sensitivity, molecular mass spectra of peptides and proteins, and provide sequence information through the process peptidide framentation.

In the proteomics section of the Central Service for Experimental Research different kinds of mass spectrometers are available, and depending on the characteristics of the sample and the information to be obtained, they determine which is the analytical technique and, therefore, the suitable mass spectrometry for each experimental application.

Sequencing of proteins

Mass spectrometry allows to obtain sequencing information through peptide fragmentation. In this way the analysis of novo sequencing can be carried out from fragmentation spectra, which allows to obtain protein microsequences of interest. 

Characterisation of proteins

Post-translational modification of proteins is a process by which living organisms regulate the function of synthesised proteins. To characterise these modifications is a complex process that can be addressed by sequence analysis by means of mass spectrometry.

Differential proteomics

The differential protein expression analysis allows the identification of proteins that change in quantity in the sample according to changes in the test environment. Knowing these changes are essential, allowing to determine biomarkers that characterise a particular cell stage. The differential expression analysis can be carried out in the proteomics section of the Central Service for Experimental Research using DIGE technology, which uses protein separation by two-dimensional gels or iTRAQ, using speed isobaric peptides present in the sample and its analysis by mass spectrometry.
In this kind of service the addressed analysis of quantification of proteins by MRM is also included.

Data analysis

The Proteomics section of the Central Service for Experimental Research has specific software for the analysis of generated data from the services offered..

Advice

A good experimental design is often essential in order to get the maximum performance of the experimental procedure chosen. Given the wide range of services offered and in order to carry out the most appropriate analyses to the type of sample and to the final objective of the experiment, the proteomics section offers scientific advice and preparation of budgeting among their services.

 
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