2022 - Nanbiosis

U30-S04. Quantitative shotgun proteomics by nLC-MS/MS

Quantitative shotgun proteomics by nLC-MS/MS

Description: Large-scale relative quantification of proteins in biological samples by nano-flow high performance liquid chromatography combined with mass spectrometry. It can use either data-dependent (DDA) or data-independent (DIA) acquisition modes. Quantification can be performed via label-free (LFQ) or protein/peptide isotopic labelling using SILAC, iTRAQ or TMT reagents. The output of this analyses are lists of proteins identified in the samples together with quantitative information obtained from the area displayed by each peptide in the mass spectra, and can then be compared across experimental conditions.

Applications: Protein profiling, biomarker discovery, drug discovery (protein target identification), development of precision medicine approaches, etc.

U30-S04 Quantitative shotgun proteomics by nLC-MS/MS

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U30-S03. Protein identification by mass spectrometry (MS) or MS/MS

Protein identification by mass spectrometry (MS) or MS/MS

Description: Identification of proteins in biological samples, either by intact protein analysis (MALDI-TOF) or protein digestion followed by peptide mass fingerprinting (PMF, MS) or peptide fragmentation (MS/MS).

Applications: Protein analysis, characterization of biological samples, analysis of protein interactions (e.g. pull-down assays).

U30 S03 Protein identification by mass spectrometry (MS) or MS/MS

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U30-S02. Determination of molecular mass of peptides, proteins and other biomolecules

Determination of molecular mass of peptides, proteins and other biomolecules.

Description: Analysis of intact proteins, peptides and other biomolecules such as oligonucleotides using mass spectrometry (MALDI-TOF or ESI-Q-TOF) for the determination of their exact molecular mass.

Applications: Characterization of biomolecules.

U30 S02 Determination of molecular mass of peptides, proteins and other biomolecules

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U30-S01. Sample preparation for proteomic analysis

Sample preparation for proteomic analysis

Description: This service includes all necessary steps to prepare samples for different proteomic analyses: protein extraction from tissues, cells or fluids, precipitation, cleaning, desalting, quantification and digestion (in gel or in solution) with proteolytic enzimes (trypsin, chymotrypsin, lys-C…), to obtain the correspondent peptides.

Applications: All experiments involving proteins.

U30 S01 Sample preparation for proteomic analysis

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U30-E05. Small equipment for protein detection/quantification

Small equipment for protein detection/quantification

Description: Under this concept, the Unit has the following instruments:

MagPix reader (Luminex), for the readout of magnetic bead–based immunoassays designed on xMAP beads. Allows the quantification of up to 50 analytes per sample.
DIGE Imager (GE), for the digitalization of difference in-gel electrophoresis (DIGE) gels or stained with fluorescence (SYPRO, etc.).
Amersham Imager 600 (GE), for the detection and digitalization of fluorescence or quimioluminiscence signals in gels or membranes.

Applications: Detection and/or quantification of proteins.

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U30-E04. Small equipment for protein separation

Small equipment for protein separation

Description: Under this concept, the Unit has the following instruments:

High-performance liquid chromatography (HPLC) system with UV detector (Agilent 1200 Series), for the chromatographic separation of proteins by reversed phase, ion exange, gel filtration or affinity.
Two IPGPhor systems (GE Healthcare), for the separation of proteins by isoelectric focusing.
Protein electrophoresis units: MiniProtean (Bio-Rad), SE600 Ruby (GE Healthcare) y Ettan™ DALTsix (GE Healthcare), for running gels up to 24×26 cm to separate proteins according to their size.
Microfraction collector and spotter SunCollect (SunChrom), to collect fractions coming out of the nLC and deposit themo onto MALDI plates for mass spectrometry.

Applications: Separation of proteins in liquid or gel supports, according to a wide variety of physicochemical properties.

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U30-E03. MALDI-TOF/TOF System

MALDI-TOF/TOF System

Description: 4800 Proteomics Analyzer (Sciex)

Technical specifications: Mass spectrometer equipped with a matrix-assisted laser desorption/ionization (MALDI) source, using a Nd:YAG 200-Hz laser, and two time-of-flight mass analyzers in tandem with a high-energy collision-induced disociation (CID) chamber for ion fragmentation. It has a variable ion path length to accomodate MS linear, MS reflector and MS/MS acquisition modes.

Applications: Determination of molecular masses of peptides, proteins and other biomolecules. Protein identification by peptide fingerprint or peptide fragmentation. Shotgun quantitative proteomics analyses by isotopic labeling and LC-MALDI workflows.

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U30-E02. QTRAP LC-MS/MS System

QTRAP LC-MS/MS System

Description: Tempo™ nanoLC-2D liquid chromatography system coupled to QTRAP5500 mass spectrometer (Sciex).

Technical specifications: The QTRAP® 5500 LC-MS/MS system (Sciex) is a triple quadrupole combined with a trap-type accelerator. It has two electrospray ionization sources (nano source and turbo source).

Applications: Determination of molecular masses of peptides, proteins and other biomolecules. Absolute quantification of proteins using isotope-labeled standards. Targeted quantitative proteomics analyses by multiple reaction monitoring (MRM).

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U30-E01. TIMS TOF Pro LC-MS/MS System

TIMS TOF Pro LC-MS/MS System

Description: Liquid nanochromatography system coupled to high resolution mass spectrometry with ion mobility (Bruker).

Technical specifications: The system consists of a nanoElute® chromatograph, which works at gradients between 50-2000 nL/min and is coupled to a TimsTOF mass spectrometer (Bruker) with an electrospray source. This spectrometer has two analyzers in hybrid configuration quadrupole and time of flight. It features a TIMS ion mobility trap that adds a fourth dimension of separation, as well as PASEF (Parallel Accumulation Serial Fragmentation) technology, which enables sequencing speeds of over 100 Hz, providing extremely high throughput and sensitivity.

Applications: Determination of molecular masses of peptides, proteins and other biomolecules. Protein identification by peptide fragmentation. Shotgun quantitative proteomics analyses by isotopic labeling (SILAC, iTRAQ, TMT…) or label-free (LFQ). Targeted quantitative proteomics analyses by PRM.

E01 TimsTOF Pro LC-MS/MS system (Bruker)

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Conversations with women scientists: Cardiology

Esther Pueyo, member of the BSICoS group, which coordinates NANBIOSIS U27 “High Performance Computing” from CIBER-BBN and I3A-UZ, participes with Lina Badimon, professor at the CSIC and group leader at the CIBERCV in the first of a series of videos where two women scientists share their experiences, prepared by the Scientific Culture and Innovation Unit (UCC+i) of the CIBER

Both researchers work in the field of Cardiology and share their passion for science and their vocation to help patients. In this video they talk in depth about their career, their difficulties, their achievements, and what their vision is about the future of women in Science and Cardiology..

“The message to university students is that they can change the future: this is what is known but you can contribute to the knowledge”`, points out Lina Badimon, who is in charge of Women in Cardiology at the European Society of Cardiology. “Women are the majority in the first stages of the scientific career but, then, women do not advance at the speed that men do and this is the point difficult to overcome for women” states Esther Pueyo.

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