The ImmunoanalysIs Core will serve as a resource to the four Projects for 1) processing, storage and sectioning of research tissues; 2) confirmation of tissue histology using routine H&E staining; 3) analysis of apoptosis using TdT-mediated dUTP-X nick end labeling (TUNEL) technique and ApopTag detection, proliferation using MIB-1 detection of Ki-67 antigen and BrdU incorporation, and androgen, glucocorticoid and estrogen receptors using monoclonal and/or polyclonal antibodies by standard descriptive immunohistochemistry and quantitative image analysis; 4) characterization of the bcl-2 family (bcl-2, bax, bcl-2, bcl-long, bak), p53 and c-myc, NFkappabeta/Ikappabeta and c/EBPbeta using immunohistochemistry; 5) demonstration of beta-galactosidase activity for cellular marking and 6) androgen receptor gene amplification and quantification of bcl-2 antisense using in situ hybridization. Drs. Mohler and Smith will co-direct the facility and supervise the activities of individuals who already possess high levels of expertise and extensive experience in performing these functions. The Core will draw upon resources currently available as well as centralize and enhance tissue processing capability through purchase of a modest quantity of equipment.
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