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Techniques

Biochemistry

  • Lipoprotein separation by sequential ultracentrifugation (Kontron)
  • Determination of blood biomarkers by autoanalizer (Cobas Mira Plus): lipid profiles, proteins, etc...
  • Determination of α-tocoferol and retinol by HPLC (Hewlett Packard, 1050 with UV/visible and fluorescence detector)
  • Apolar aldehide quantification by HPLC (Hewlett Packard, 1050 with UV/visible and fluorescence detector)
  • Total aldehide quantification by TLC
  • Determination of the susceptibility to oxidation of lipoproteins by spectrophotometry (Uvikon 922, Kontron)
  • ELISAs: TNF-a, FABP4, ApoH, E-selectine, VCAM-1, ICAM-1, ox-LDL, Ab-oxLDL ...

Cellular cultures

  • Optimization and mantainence of primary cultures and stable cellular lines
  • Primary cultures: fibroblasts, monocytes, HUVEC, HAEC, VSMC ...
  • Cellular lines: THP-1, HEPG2, Jurkat, 3T3L1 ...
  • Freezing and thawing of cells.
  • Storage of cells in liquid nitrogen.
  • Counting of cells with a Neubauer camara (Olympus microscope, inverted and phase contrast).
  • Determination of the cellular viability with Trypan blue, LDH, Hoechst.
  • Detection of micoplasms in cellular cultures by PCR.
  • Specific cellular colouring: Oil Red, ORO ...
  • Cellular adhesion assay.


Cellular biology

  • Obtaining cellular nuclear protein extracts.
  • Obtaining cellular lisats.
  • Protein determination by Lowry, Bradford
  • Protein Separation by electrophoresis with poliacrilamide gels (SDS-PAGE ...) and silver nitrate colouring, coomassie blue (Protean II, BioRad; X Cell SureLock Mini Cell, Invitrogen; Multiphor II System, Amersham Biosciences; Hoefer SE 260, Amersham Bioscience)
  • Protein determination by western blot: XCell Blot Module (Invitrogen) Trans-Blot SD Semi-Dry Transfer Cell (BioRad)
  • Immunoprecipitation


Molecular Biology

  • Total RNA Isolation from tissues and cells.
  • cDNA synthesis. inversa Transcripció (RT)
  • Arrays of genetic expression Arrays (in collaboration with an outside company)
  • DNA amplification by PCR
  • Electrophoresis with poliacrilamide gels and silver nitrate colouring, Sybr-Safe.
  • Electrophoresis with agarose gels and with Sybr-Safe colouring.
  • Electromobility shift assay (EMSA)
  • Cloning of promotors in vectors
  • Directed mutagenesis
  • Cellular transfeccions
  • Luciferase assay

Staff in the URLA laboratories work according to ISO 9001 quality control.


IISPV

IISPVInstitut d'Investigació Sanitària Pere Virgili (IISPV)

Work Center for R&D&I in Nutrition and Health
Avda. de la Universitat, 1 - Second floor
E-43204 Reus (Tarragona)

Tel. Work+34 977 75 93 94 http://www.iispv.cathttp://www.iispv.cat

Generalitat de Catalunya

Universitat Rovira i Virgili (URV)

ICS - Institut Català de la Salut

Grup Sagessa

Centre de Recerca de Catalunya

Institut Pere Mata, Hospital Psiquiàtric Universitari

Spain Excellence University - Universitat Rovira i Virgili, Excellence Research Spain, Excellence Postgraduate Spain

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