The purpose of Core C is to provide, on request, molecular analysis services to the investigators in this Program Project. The services will include Western immunoblot;in situ hybridization;quantitative (real-time) reverse-transcriptase polymerase chain reaction (qRT-PCR) analyses;electrophoretic mobility shift assays; and cytokine bioactivity analyses. In addition, analyses of other proteins and their encoding mRNAs that may be of interest (e.g., other cytokines, growth factors, or structural proteins) will be available upon request. We will have A(3, S100B, and GFAP ELISAs available for rapid screening of large numbers of samples. As a rapid screen for specific mRNAs in cell cultures, micro-//? situ hybridization will also be available. The tissue to be analyzed will be from human and rodent brain (Projects 1 and 2) and from cell cultures (Projects 1 and 3). A key aspect of the Core will be provision of expertise in time-consuming steps, such as antibody selection and optimization of PCR primer design. In addition, standardization across projects will help to achieve valid comparisons and improve efficiency. Results from analyses performed in this Core will be provided to Project Leaders primarily as raw data, including appropriate controls to document sample integrity and equipment function. Data for some analyses will be supplemented by routine extrapolation, and Core personnel will consult with Project Leaders in developing sophisticated parsing of data and experimental parameters. Residual samples and processed specimens, such as tissue sections, will be returned to the respective Project Leader for archiving. Records of services performed and quantitative data from the Core will be archived in the Core's computer database, under protection of state-of-the-art firewalls and data backup. Achieving the goals of this Core will allow standardization of methods for data acquisition and analyses across projects and will help ensure efficient collection and dissemination of data for publication.
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