OPTIMIZATION OF THE IMMUNOHISTOCHEMICAL DEMONSTRATION OF KERATINS IN PARAFFIN WAX-EMBEDDED MOUSE SKIN

The purpose of this study was to develop an immunoperoxidase technique for the detection of cytokeratins in samples of paraffin wax-embedded adult and fetal skin from NMRI mice, with various antibodies (Troma-1 , LL001, 8.60, MCK5, MCK6, AF129) that have been tested mainly on fres h-frozen sections. Each antibody was tested with three different fixat ives (10% neutral buffered formalin, Bouin's fluid, and 70% ethanol) a nd two distinct pretreatments (enzymatic digestion with trypsin, or he at treatment). The best results, in terms of non-specific background l abelling, morphological preservation and intensity of specific labelli ng, were obtained (1) for adult skin, by the use of Bouin's fluid, hea t pretreatment and antibodies LL001, MCK5, MCK6 or AF129, and (2) for fetal skin, by the use of 70% ethanol, heat pretreatment and antibody Troma-1. Monoclonal antibody 8.60 gave the best results when the use o f 70% ethanol was combined with either enzymatic digestion or heat pre treatment. (C) 1998 W.B. Saunders Company Limited.