CORE--MOLECULAR ANALYSIS

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The purpose of Core C is to provide molecular analysis services to the Project investigators in this Program Project. Western immunoblot, northern and in situ hybridization analyses (to determine the levels of IL-1, S100beta, and beta-APP and their encoding mRNAs, as well as other proteins that may be of interest, e.g., TGF-beta, and FGF) will be available upon request; S100beta and GFAP ELISAs will also be available for rapid screening of large numbers of samples. The levels of total protein, total RNA, and ribosomal RNA will be performed as routine analyses necessary for determining the relative levels of the specific proteins and mRNAs mentioned. The human tissue to be analyzed for studies in Project 1 will have been obtained by Project investigators from the Neuropathology Core (B1 and B2). The animal tissue or cells will be from rats used in experiments in Projects 2 and 3. Data from analyses performed in this Core will be provided to the Project Investigators as both raw data and preliminarily analyzed data. For example, data from northern and western immunoblot analyses will be returned to the investigator as raw data (pictures of gels and films of transfers) and ng/micrograms tissue or ng/micrograms of extract. Data from ELISAs will be returned as raw data in the form of optical density units per sample, together with a standard curve generated at the same time, and calculation of ng/micrograms protein and ng/micrograms extract. Following in situ hybridization, sections will be returned to the investigator or sent to Core D for analysis. Data from in situ hybridization of cultures will be returned to the investigator as cpm/well. Values from studies of human and animal tissue will be entered into the relevant database.