The Biochemical Core Laboratory of this Program Project Grant will provide a consolidated highly specialized well-equipped and professional unit capable of performing a variety of radioimmunoassay, biochemical and molecular procedures as required by project investigators. No single investigator laboratory supported by R01 would be capable of equipping and staffing this resource. The Core will carry out all the analytical techniques: 1) Assay of CYP and NOS activities by radiochemical HPLC. 2) Measurement of 20- HETE and EETs in tissue samples, CSF and microdialysates by LC/MS. 3) Identification of novel metabolites generated by CYP4F and CYP4X enzymes by LC?MS/MS. This Core is also equipped to measure IP3, DAG, microglial and astroglial products in the brain and cerebral vasculature. In addition we will measure CYP and NOS protein using western blots and immunohistochemistry. Molecular measurements with real time RT-PCR and in situ hybridizations will be carried out as outlined in the projects. Access to the analytical expertise and equipment in the central facility is essential to each of the proposed projects and justified by the large number of biochemical and molecular determinations required for successful completion of the proposed studies.
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