A murine monoclonal antibody specific for calf intestinal alkaline phosphatase has been prepared and used in an unlabeled antibody bridge technique for labeling monoclonal antibodies. This procedure-the alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) method-gives excellent immunocytochemical labeling of tissue sections and cell smears, comparable in clarity and intensity to that achieved with immunoperoxidase labeling. If the enzyme label is developed with a naphthol salt as a coupling agent and Fast Red or hexazotized new fuchsin as a capture agent, a vivid red reaction product is obtamed which is very easily detected by the human eye. For this reason the APAAP technique was found particularly suitable for labeling cell smears (for both cytoplasmic and surface-membrane antigens) and for detecting low num-APAAP 220 CORDELL ET AL APAAP STAINING OF MONOCLONAL ANTIBODIES

The p53 gene mapped on chromosome 17p is an important negative regulator of normal cell growth and division (Finlay et al., 1989;Chen et al., 1990;Diller et al., 1990;Isaacs et at., 1991). Alteration or inactivation of p53 by mutation can allow a cell to escape from normal into uncontrolled growth leading to cancer development. p53 mutations are a very common genetic change in a variety of human tumours Takahashi et al., 1989;Nigro et al., 1989;Bartek et al., 1990a;Marks et al., 1991;Tamura et al., 1991). The majority of the mutations alter the conformation of the nuclear protein product, encoded by the p53 gene (Gannon et al., 1990;Bartek et al., 1990b; Rodrigues et al., 1990). In normal cells and tissues the p53 protein has a very short half-life (Oren et al., 1981) and attains such a low level that it is not detectable histologically (Gannon et al., 1990). The mutant forms have an extended half-life (Finlay et al., 1988) and being overexpressed are readily detected by immunohistochemistry. Elevated p53 levels have been described in different human neoplasms (Crawford et al., 1984;Cattoretti et al., 1988;Bartek et al., 1990a;Iggo et al., 1990;Davidoff et al., 1991a;Marks et al., 1991

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SUMMARY A panel of seven monoclonal antibodies was applied to smears of cell deposit from 70 pleural and peritoneal fluids, using an immunoalkaline phosphatase (IAP) procedure. The cases were chosen to show typical cytological patterns, both benign and malignant, and in this way the diagnostic value of the method could be assessed.The antibodies used were 2D1 (anti-leucocyte), Ca 1, HMFG-2 (anti-milk fat globule membrane), LE61

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