Proteomics Unit

The Proteomics Unit offers researchers various services relating to the field of proteomics, as well as seminars and training courses on this matter.

The Proteomics Laboratory, on the other hand, provides a diverse range of protein analysis services, many of which require the contracting of external services. In addition, most of these services entail the need for a close technical-scientific interaction with the user in order to determine the best analytical strategy to use.

The priority of these analyses will be given, in all cases, to the samples sent by research groups and other academic institutions belonging to the i+12 Institute. All analyses requested by external users (other research bodies and private companies) will be assessed by the Unit Manager.

The Proteomics Unit supports and services the following technologies:

1. Protein fractioning and differential expression analysis.
   1. 2D electrophoresis for the conventional analysis of gel protein mixtures.
   2. 2D electrophoresis for quantitative differential analysis using the “DIGE” technology.
   3. Multiple isobaric labeling.
2. Protein identification in gel and solution.
3. Separation and analysis of Protein Complexes under native conditions: BLUE NATIVE. This analysis is performed both in 1D and 2D, and involves the transfer to membranes or the staining of gels for subsequent protein identification.
4. Theoretical-practical assistance on the specific design of each type of immunoblotting technique.
5. Unidimensional differential expression analysis.

To implement these techniques and methods, the Proteomics Unit currently features the following high-tech equipment:

EttanTM 2D-PAGE Bidimensional Electrophoresis System

• This system contains an IPGphor™ III unit that makes up the isoelectric focusing system used to perform the first-dimension electrophoresis. It enables the use of immobilized pH gradient gel strips using the IPGphor™ Manifold tank that allows for simultaneously running up to 12 strips. The IPGphor™ III unit includes a control software that generates graphs where voltage, current and volt-hour fluctuations taking place during the isoelectric focusing program are monitored.
• The DALTsic electrophoresis unit, where the second-dimension is performed, consists of an electrophoresis chamber that allows for running up to six gels at a time, thus reducing the experimental variability in each of them. It also features a gel cassette carrier designed to polymerize up to six gels at once.
• Both units are stabilized by the MultiTemp ™ III (GE Healthcare) system, which offers a reliable and reproducible temperature control system by circulating liquids at a constant temperature through the external electrophoresis devices. Its great cooling capacity (200 V) allows for connecting up to two vertical electrophoresis units at the same time in order to achieve a greater assay reproducibility.

3100 OFFGEL (Agilent) Liquid-Phase Isoelectric Focusing Fractionator

• Preparative liquid-phase isoelectric focusing: the Offgel Agilen 3100 fractionator separates complex protein or peptide blends according to their isoelectric points (Ip), thus obtaining high-resolution fractions. The resulting fractions stay in the solution, thereby facilitating their recovery for LC/MS analysis or analyses with conventional gels. The results of this technique are highly reproducible.

2100 Bioanalyzer (Agilent Technologies)

• This system is based on microfluidic capillary electrophoresis; therefore, it can determine, quantify and demonstrate the quality of total protein extracts.

ImageQuant LAS 4000 (GE Healthcare) gel analysis and documentation system

• This is an automated image analysis system that combines an optimal capture and analysis of images using the ImageQuantTL software. The system uses a 3.2 megapixel CCD camera that allows for capturing digital images of gels and chemiluminescent, stained or fluorescent membranes with an effective resolution of 6.3 megapixels.

ChemiDoc MP Imaging System (Bio Rad) for gel analysis and documentation

• Imaging system used to cover multiplex fluorescence, chemilomuniscent and other general gel documentation applications using a CCD camera.

Gel DocTM EZ (Bio Rad) gel analysis and documentation system

• This is a compact and automated image analysis sytem that combines an optimal capture and analysis of images using the Image Lab software. It allows for documenting mini-gels stained with fluorescent or visible dyes, photographic plates and stain-free gels (Bio-Rad stain-free).

ImageScanner III (GE Healthcare)

• A high-resolution scanner used for the digitalization and densitometric quantification of gels with visible stains.

Maxwell 16 (Promega) nucleic acid purification system

• Purification system for proteins with polyhistidine tails that decreases the time needed to purify this type of tagged proteins and processes a greater number of samples at the same time.

The head of this Unit is Dr. Inés García-Consuegra Galiana, Senior Technician in Research Support. The Unit is located in Plant 0 of the Research Center of the Hospital Universitario 12 de Octubre.

Contact information:

• e-mail: inesgcg@h12o.es
• Telephone No.: 91 390 8000/ Ext.: 1465
• Fax No.: 91 390 8455

Access to rates of the Proteomics Unit.