Dorothy Hodgkin Proteomics and Biochemistry

1. SERVICE DEFINITION

2. SERVICES OFFERED

• Sample analysis by SDS-PAGE electrophoresis (1 or 2 dimensions)
• Western Blotting assays (protein detection and quantification)
• Differential gel electrophoresis (DIGE) assays and analysis
• Purification of recombinant proteins on demand
• Set-up of protein production
• Study of molecular interactions (SPR or ITC)
• Characterization of enzymatic activities
• Technical assistance in the design and execution of experiments with proteins
• Protein quantification
• Proteome analysis (with or without enrichment)

3. PERSONNEL AND CONTACT

Facility Director

            Dra Laura Tomás Gallardo - 954978208

Associated technicians:

            Cynthia Alias Villegas

4. EQUIPMENT

•    Sistema de imagen ChemiDoc MP
•    Espectrofotómetro UV-Vis Shimadzu UV-1280
•    Escáner Image scanner III (Cytiva)
•    Escáner Láser Amersham Typhoon RGB (Cytiva)
•    Sistemas de electroforesis
•    IPGPhor (Cytiva)
•    Sonicador Branson SFX 550
•    Biacore X100 (Cytiva)
•    Nanocalorímetro PEAQ-ITC (MicroCal - Malvern)
•    Cabina de flujo laminar Cruma HZ-2
•    Microcentrífuga 5415R (Eppendorf)
•    Incubado Innova 42 (New Brunswick- Eppendorf)
•    Sistema de transferencia Trans-Blot Turbo (BioRad)
•    ÄKTA™ Pure 25 FPLC (Cytiva)
•    Liofilizador Telstar LyoQuest -85

5. RATES

¹ SOLICITAR PRESUPUESTO SEGÚN CARACTERÍSTICAS DEL EXPERIMENTO
² Incluye, lisis, inhibidores de proteasas, limpieza y cuantificación.
³ Incluye electroforesis, transferencia, detección y análisis de imagen. El usuario aportará el anticuerpo.   
⁴ Incluye tinción del gel.

6. PUBLICATIONS AND WEB RESOURCES

Publications:

• Fine-Tuning Modulation of Oxidation-Mediated Posttranslational Control of Bradyrhizobium diazoefficiens FixK(2) Transcription Factor. Int. J. Mol. Sci. 2022, 23, 5117. https://doi.org/10.3390/ijms23095117
• Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos. STAR Protocols, 2022, 3, 101058. https://doi.org/10.1016/j.xpro.2021.101058
• Dissection of FixK(2) protein-DNA interaction unveils new insights into Bradyrhizobium diazoefficiens lifestyles control. Environmental Microbiology, 2021, 23, 6194. https://doi.org/10.1111/1462-2920.15661
• CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos. Developmental Cell, 2020, 54, 805-817. https://doi.org/10.1016/j.devcel.2020.07.013
• The CbrB Regulon: Promoter dissection reveals novel insights into the CbrAB expression network in Pseudomonas putida. PLOS ONE, 2018, 13, e0209191. https://doi.org/10.1371/journal.pone.0209191
• Searching for a Cell-Based Therapeutic Tool for Haemophilia A within the Embryonic/Foetal Liver and the Aorta-Gonads-Mesonephros Region. Thromb. Haemost., 2018, 118, 1370. https://doi.org/10.1055/s-0038-1661351

Web resources:

• Transcription and Translation Tool: Converts sequences from DNA to RNA and from this to protein.

           http://biomodel.uah.es/en/lab/cybertory/analysis/trans.htm

• ProtParam tool: allows the computation of various physical and chemical parameters for a given protein.

            https://web.expasy.org/protparam/

• SnapGene Viewer: View, annotate and share plasmid sequence files.

            https://www.snapgene.com/snapgene-viewer

• Expasy: Swiss Bioinformatics Resource Portal

            https://www.expasy.org/

• Uniprot: Base de datos de secuencias de proteínas e información funcional.

            https://www.uniprot.org/

• Fold-It: Programa de ciencia ciudadana que permite contribuir a investigaciones científicas mediante una dinámica lúdica de plegamiento de proteínas.

            https://fold.it/

• AlphaFold: Base de datos de estructuras de proteínas resueltas por IA.

            https://alphafold.ebi.ac.uk/