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analyTical techniques

Laboratory Groups Biomedical Repository Brain Function & Dementia Group Cytokines and Inflammation Group Infection & Immunity Group Vascular Research Group

Quantitative techniques for analysing genes and proteins in human samples are key to understanding mechanisms of human disease and for monitoring disease progression and response to treatment. SRBF researchers have expertise in a range of techniques and are willing to provide advice to Trust researchers who wish to apply these to clinical research questions.  Immunoassay The unit undertakes a variety of specialised immunoassays for cytokines and other disease markers using standard enzyme linked immunosorbant assays (ELISA) and multiplex analysis using Luminex® technology. Specialist expertise in development and standardisation of new immunoassays for research studies is available.  Contacts:   Steve Hopkins, Margaret Hoadley (ELISA and Luminex assays) Steve Hopkins, Rob Oliver (Immunoassay development) Gene Expression The unit has excellent facilities for analysis of human gene expression and small-scale genotyping using Lightcyler480® real-time PCR. Expertise in knockdown of genes in human cell lines using siRNA technology Contacts:   Geoff Warhurst (PCR and gene suppression) Steve Hopkins, Margaret Hoadley (Cytokine PCR) Molecular Microbiology PCR-based assays for the detection and species identification of microbial DNA in blood and other clinical samples as tools to assist in infection diagnosis Contacts: Geoff Warhurst Human Cell Culture Expertise in isolation and primary culture of human cells (leucocytes; vascular endothelial and smooth muscle cells) and cell lines (gut epithelial, human hepatoma HepG2 cells) Contacts: Kirk Siddals (Vascular Smooth Muscle; Vascular Endothelial; HepG2) Steve Hopkins (Leucocytes and whole blood) Geoff Warhurst (Gut epithelial; Caco-2, HT29) Isolated Tissues Techniques for analysis of gut permeability and drug transport using ex vivo human tissues mounted in Ussing chambers. Contacts: Geoff Warhurst, Norman Higgs Electrophoretic analysis Expertise in the analysis of proteins by native, denaturing and reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and 2D PAGE. We have extensive experience in the development and optimisation of western immunoblotting. Contact: Kirk Siddals Chromatography Expertise in the analysis and preparative purification of biomolecules by high performance liquid chromatography (HPLC) and column chromatography. Specific techniques include size exclusion chromatography, reverse phase HPLC and ion-exchange HPLC. Contact: Kirk Siddals Histology Extensive experience of preparing brain tissue for histochemical and immunohistochemical analysis. Contact: David Mann