- Related scientific areas
- Biología y Biomedicina
- Ciencia y Tecnología de Alimentos
- Ciencias Agrarias
- Group
- Proteómica, Genómica y Metabolómica
- Subgroup
- Proteómica
Molecular weight analysis by MS-MALDI TOF
Service general data
Service data
- Support unit: PROTEÓMICA Y GENÓMICA
- Institute: CENTRO DE INVESTIGACIONES BIOLOGICAS MARGARITA SALAS
- Locality: Madrid (Madrid)
- Service's web: http://www.cib.csic.es/es/servicio.php?iddepartamento=29
Service's description
PROTEÓMICA El equipamiento actual del Servicio es el siguiente: 2 unidades de isoelectroenfoque Protean IEF cell (BIO-RAD), cubeta multigel Miniprotean Dodecacell, cubeta Protean II Multicell, programa de análisis y cuantificación de imágenes PDQuest, EXQuest Spot Cutter (BIO-RAD), digestor de proteínas automático INTAVIS, espectrómetro de masas de MALDI-TOF-TOF Autoflex III (Bruker) y espectrómetro de masas LTQ Orbitrap Velos (Thermo Scientific) dotado de una fuente de ionización nano (Proxeon) conectada "on-line" a un nano-HPLC Easy-LC (Proxeon). GENÓMICA Equipos disponibles: Nanodrop ND-1000, 2 sistemas de electroforesis en chip Experion (BIO-RAD), sistema iQ5 (BIO-RAD) de PCR cuantitativa en tiempo real (rt-PCR) (disponemos de la colección de sondas Taq Man de genoma humano Universal Probe Library de ROCHE).
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Molecular weight analysis by MS-MALDI TOF
Service's description
Proteins
Peptides
Glycoproteins and glycopeptides
Oligosaccharides
Oligonucleotides
Natural or synthetic polymers
Synthesis products
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- No hay prestaciones de servicio asociadas a esta unidad
- Peptide Mass Fingerprinting MS - MS/MS by MALDI TOF/TOF
- Affinity chromatography: Enrichment in phosphopeptides for proteomics
- 2D Electrophoresis
- 1D Electrophoresis SDS-PAGE
- Protein Identification by LC/MS/MS (>50 proteins)
- Nanodrop: Spectrophotometry of liquid samples.
- Identification and Characterization - Real Time PCR
- RNA quality analysis
Service general data
Service data
- Support unit: PROTEÓMICA
- Institute: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA
- Locality: Madrid (Madrid)
- Service's web: http://www.cbm.uam.es/proteomica.htm ,http://www.proteored.org
Service's description
Desde 1993 la trayectoria del Servicio de Proteómica del CBMSO ha sido paralela a la evolución de la comunidad científica en el estudio de las proteínas y sus objetivos son ofrecer apoyo y asesoramiento al investigador con métodos apropiados para preparar la muestra y diseñando el flujo de trabajo óptimo para cada objetivo. El laboratorio está equipado con sistemas de electroforesis 1D y 2D y tres espectrómetros de masas, un MALDI-TOF (Bruker), un LTQ-VELOS (2D-Trampa iónica en tándem-Thermo-Scientific) y un LTQ-ORBITRAP-VELOS-PRO (2D-Trampa iónica en tándem acoplada a un analizador Orbitrap-Thermo-Scientific). Nuestro Servicio ha sido usado por numerosos proyectos de investigación en toda España. Servicios: Análisis de Peso Molecular de Péptidos y Proteínas Preparación de Muestras Identificación de Proteínas mediante Mapeo Peptídico (MS-MALDI-TOF) Identificación de proteínas mediante LC-MS/MS (Ion Trap) Caracterización de Proteinas Secuenciación De Novo
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Molecular weight analysis by MS-MALDI TOF
Service's description
Matrix Assisted Laser Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) allows analysis of proteins and peptides with high sensitivity and soft ionization with little or no fragmentation. Ions are formed through gas-phase proton-transfer reactions with photoionized matrix molecules. Matrix is typically a small energy absorbing molecule such as 2, 5-dihydroxybenzoic acid (for synthetic peptides) or sinapinic acid (for intact proteins). MALDI-TOF MS analysis produces data for peptide and protein molecular weight determination.
MALDI-MS is also sensitive to the presence of contaminants such as salts or surfactants, and protein-containing solutions that have high levels of these contaminants often exhibit little or no protein signal. Such contamination intrinsically results from some purification procedures, and in certain cases, salts or surfactants are required to keep proteins from precipitating from solution. Contaminants likely reduce MALDI-MS signals by disrupting the cocrystallization of analytes and matrix molecules.
To avoid this problem we recommend:
1. Dyalisis of the sample.
2. Submit the sample as concentrated as possible to allow us to be diluted in the appropriate solvent.
3. Desalt the sample by zip-tip, oasis, omix¿(pipette tips/columns containing C18 or C4 media)
The amount needed for the analysis of peptides is the range of femtomoles and for proteins is the range of picomoles.
Procedure:
An aliquot (0.5 ul) of the sample (protein/peptide) is analyzed by a MALDI-TOF-type of mass spectrometry using an Autoflex model of Bruker Daltonic (Bremen, Germany), in a positive ion reflector mode, and employing (0.5 ul) sinapinic acid for proteins and 2,5-dihydroxybenzoic acid (0.5 ul) for peptides as matrix and an Anchor-Chip surface target (Bruker Daltonic). Peak identification and monoisotopic peptide mass assignation were performed automatically using Flexanalysis¿software, version 2.2 (Bruker Daltonics).
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- No hay prestaciones de servicio asociadas a esta unidad
- Protein Identification by LC-MS/MS (<50 proteins)
- Protein Identification by LC-MS/MS (<5 Proteins)
- Protein Identification by LC/MS/MS (>50 proteins)
- Peptide Mass Fingerprinting by MS-MALDI TOF
- Coomassie Gel Staining
- Data analysis, experimental design, training and advise
- Sample preparation
- 1D Electrophoresis SDS-PAGE
- Determination of post-translational modifications
- Desalting and concentration by Zip-Tip
Service general data
Service data
- Support unit: SERVICIO DE PROTEÓMICA IIBB
- Institute: INSTITUTO DE INVESTIGACIONES BIOMEDICAS DE BARCELONA
- Locality: Barcelona (Barcelona)
- Service's web: proteomica.uab.cat
Service's description
Laboratorio de Análisis Proteómico provisto de la tecnología necesaria para el análisis cualitativo y cuantitativo de proteomas a gran escala así como para la identificación y cuantificación de péptidos y proteínas diana. El laboratorio ocupa un espacio de 90 m2 en el que se disponen 4 espectrómetros de masas (4800 MALDI-TOF/TOF, Quantum Triple Quadrupole , LTQ Velos Linear ion trap y Orbitrap XL), diversos sistemas acoplados para cromatografía líquida convencional y nanocapilar (2xAgilet 1200, 2X Agilent 1100) y un sistema de IEF preparativo (IEF off-gel). Dispone asimismo de un robot para digestión automática (DigestPro MS, Intavis) y de los sistemas necesarios para la realización de geles 2D de tamaño pequeño y grande (2 x IPGphor y 2 x Ettan Dalt VI (GE), Protean y MiniProtean (BioRad)) así como para la adquisición y análisis de las imágenes (Scanner y SameSpots software). Dispone asimismo de aparataje propio necesario para el trabajo diario (centrífugas, arcones, etc).
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Molecular weight analysis by MS-MALDI TOF
Service's description
Fairly complex peptide or protein mixtures can be analyzed without previous separation (The technique can be used to obtain proteome fingerprints which is the basis of protein profiling technologies such as SELDI).
Samples must be delivered dissolved in a buffer, preferably a volatile one, with a salt concentration lower than 50 mM.
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- Proteomic quantitative analysis by TMT
- Data analysis, experimental design, training and advise
- Coomassie Gel Staining
- FASP digestion
- Determination of post-translational modifications
- Protein identification by de novo sequencing
- Protein Identification by LC-MS/MS (<50 proteins)
- Protein Identification by LC-MS/MS (<5 Proteins)
- Peptide Mass Fingerprinting MS - MS/MS by MALDI TOF/TOF
- 1D Electrophoresis SDS-PAGE
- Protein Identification by LC/MS/MS (>50 proteins)
- Peptide Mass Fingerprinting by MS-MALDI TOF
- Protein Separation by HPLC
- Protein Identificacion by 2D LC-MS/MS (>5 fractions)
- Protein Identification by 2D LC-MS/MS (2 - 5 fractions)
- Silver Gel Staining
Service general data
Service data
- Support unit: UNIDAD DE PROTEÓMICA
- Institute: INSTITUTO DE PARASITOLOGIA Y BIOMEDICINA LOPEZ NEYRA
- Locality: Armilla (Granada)
- Service's web: http://www.ipb.csic.es/servicios/Proteomica/index.html
Service's description
El principal objetivo de la Unidad es proporcionar soporte tecnológico en el análisis de proteínas y otras biomoléculas, mediante el uso de la electroforesis, la cromatografía y la espectrometría de masas. Las diferentes prestaciones se ofrecen a Investigadores de Centros y Entidades de titularidad tanto pública como privada. Entre estas, destacar: - Determinación de masa molecular de biomoléculas - Identificación y caracterización de proteínas mediante huella peptídica y fragmentación, con MALDI-TOF/TOF y con nLC-MS/MS - Cromatografía líquida en columna Equipamiento: - Espectrómetro de masas MALDI-TOF/TOF UltrafleXtreme (Bruker) - Espectrómetro de masas ESI-Ion Trap (IT) Amazon Speed ETD (Bruker) - Cromatógrafo Easy-nLC II (Thermo) - Colector de fracciones LC-MALDI Proteineer fc II (Bruker) - Picador de geles ExQuest Spot Cutter (Bio-Rad) - Digestor automático DigestPro MS (Intavis AG) - FPLC AKTA Purifier (GE Healthcare) - Equipo electroforesis en gel (Bio-Rad)
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Molecular weight analysis by MS-MALDI TOF
Service's description
This type of analysis is carried out from samples in solution at known concentrations (or lyophilized samples in estimated quantities). Samples can be proteins, peptides, DNA and RNA oligonucleotides, PNAs and other types of biological or synthetic polymers. Depending on the type of sample, different matrices are used that co-crystallize with it.
The analysis includes sample dilution to the optimum concentration for analysis, acquisition of mass spectra in the specified mass range and a report of results in which the procedure followed is indicated and the obtained spectra, mass list and intensities are included.
It is recommended that the samples be dialyzed or dissolved in a volatile buffer and as concentrated as possible, so that they can be diluted with suitable solvents. In the case of samples containing components (surfactants, salts, etc.) that interfere with the crystallization process, the corresponding variant will be selected and desalination and concentration will be carried out to eliminate the interferences.
For the analysis of proteins and oligonucleotides, quantities of the picomoles/ul range and in the case of peptides, femtomoles/ul are required. A minimum volume of about 5 ul will be required. The larger the size of the molecules to be analyzed, the greater the requirement for sample quantity.
For molecules between 500 Da and 5 KDa, analysis will be carried out in reflector mode, with an estimated error of 10 ppm. For molecules larger than 5KDa, linear mode will be used, whose estimated error is 0.01% (100 ppm).
INSTRUCTIONS FOR SUBMITTING SAMPLES
1.- The sample in solution or lyophilized must be sent in a 0.5 ml tube clearly identified with the sample name labeled on the lid.
2.- Samples will be delivered or sent along with the corresponding application properly filled out (MOLECULAR WEIGHT DETERMINATION AND FRAGMENTATION REQUEST), where the concentration and solvent composition (protein quantity of dried or lyophilized samples) must be specified.
3.- It is important to take into account the different contaminants that can interfere in the process of analysis by mass spectrometry (INTERFERENTS).
NOTES:
If it is a protein sample in gel, it must be previously extracted from it.
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- No hay prestaciones de servicio asociadas a esta unidad
- Desalting and concentration
- Molecular weight analysis by Mass Spectrometry: ESI by MS-QTOF, MS-ION TRAP, Orbitrap-MS, FT-ICR-MS or TSQ-MS
- Protein Separation by HPLC
- Protein quantification
- Sample filtration
- Technical and Experimental Consulting
- Protein identification using LC-ESI-MS/MS
- Additional reagents
Service general data
Service data
- Support unit: SERVICIO DE ANÁLISIS DE PROTEÍNAS
- Institute: INSTITUTO DE BIOQUIMICA VEGETAL Y FOTOSINTESIS
- Locality: Sevilla (Sevilla)
- Service's web: http://www.ibvf.csic.es/descripcion-analisis-proteinas
Service's description
El objetivo principal del Servicio de Análisis de Proteínas del IBVF es la identificación de proteínas de interés biológico, así como la determinación de masas moleculares de péptidos y proteínas en solución, mediante espectrometría de masas. Para ello, cuenta con diferentes equipos para la preparación y el análisis de las muestras: -Sistema para el recorte automatizado de proteínas en gel de una o dos dimensiones, Modelo EXQuest Biorad. -Espectrómetro de masas con analizador de tiempo de vuelo e ionización por desorción láser asistida por matriz (MALDI-TOF) con capacidad para fragmentar péptidos, Modelo 5800, ABsciex. -Espectrómetro de masas híbrido, Triple cuadrupolo-TOF acoplado a nanoLC, Modelo 5600, ABsciex que permite análisis LC-MS/MS.
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Molecular weight analysis by MS-MALDI TOF
Service's description
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- No hay prestaciones de servicio asociadas a esta unidad
- Protein Identification by LC-MS/MS (<50 proteins)
- Protein Identification by LC-MS/MS (<5 Proteins)
- Protein Identification by LC/MS/MS (>50 proteins)
- Peptide Mass Fingerprinting by MS-MALDI TOF
Service general data
Service data
- Support unit: PROTEÓMICA
- Institute: INSTITUTO DE QUIMICA AVANZADA DE CATALUÑA
- Locality: Barcelona (Barcelona)
Service's description
El objetivo principal del servicio es dar apoyo científico y tecnológico en el área de la proteómica a los investigadores del IQAC y IDAEA así como de otros ámbitos, tanto públicos como privados. Entre nuestras principales actividades se encuentra la separación, cuantificación, identificación y caracterización de péptidos y proteínas en sistemas de interés biológico y biomédico mediante técnicas de electroforesis bidimensional y espectrometría de masas. Otras actividades que realizamos mediante espectrometría de masas es la determinación de la masa molecular de diferentes especies moleculares tanto de origen sintético como ambiental facilitando así su elucidación estructural. El servicio dispone de la técnica MALDI MSImaging que permite detectar la distribución de moléculas de diferentes masa en un corte de tejido u otro tipo de material sin necesidad de marcaje. El laboratorio dispone de un sistema completo de separación de proteínas mediante electroforesis bidimensional.
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Molecular weight analysis by MS-MALDI TOF
Service's description
Samples must be in solution. Depending on the molecular mass (small 20,000 Da), the concentration necessary for the analysis could vary from few femtomoles to hundreds of picomoles. The solution should be free of salts, detergents and other contaminants which may impede or hinder the crystallization of the sample with the matrix.
In the case of special samples, and if the user agrees, studies with different solvents and matrix types will be made in order to optimize the results.
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- No hay prestaciones de servicio asociadas a esta unidad
- Peptide Mass Fingerprinting MS - MS/MS by MALDI TOF/TOF
- 2D Electrophoresis
- 1D Electrophoresis SDS-PAGE
- Peptide Mass Fingerprinting by MS-MALDI TOF