CORE--AUTOMATED COMBINED IN SITU HYBRIDIZATION AND IMMUNOHISTOCHEMISTRY

University Of Arizona, Tucson AZ

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Abstract

CORE COMPONENT C: Automated combined in situ hybridization (ISH) and immunohistochemistry (IHC) laboratory. The purpose of this core is to provide a facility for automated in situ hybridization and immunohistochemistry which will specifically assist in the successful completion of Projects 1 and 3. Using instrumentation developed at the University of Arizona, the Core Leaders have developed a kinetic-mode automated assay which facilitates rapid combined ISH and IHC assays. With this procedure, reactions can be performed within a 1-3 hour time frame (1). Double labeling IHC and ISH are of benefit: (a) in co- localizing interactive molecules relevant to pathogenesis; (b) in relating RNA message to protein product; (c) in demonstrating unique diagnostic immunoarchitectural features; and (d) in demonstrating clonality of tumor cells. This core will directly impact Project 1 (Specific Aim #1) wherein the question of the importance of alpha3 and alpha6 integrin expression will be examined. It will also aid in Aim 2 where the effect of laminin 5 on hemidesmosome formation will be examined. Core C will support Project 2 in Specific Aims 1 and 2 in which the effect of beta 4 and alpha 6t transfection on invasion will be tested. Core C will support Project 3 by providing both protein and mRNA localization to study the expression pattern of matrilysin in subcutaneous and orthotopic LNCaP tumors in immunodeficient mice.