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.