Maspin (mammary serine protease inhibitor), a member of the serpin family, has been shown to inhibit angiogenesis, tumor invasion, and metastasis. Previous studies suggest a p53-dependent regulatory pathway of maspin protein expression. Its loss correlates with progression of disease in both breast and prostate cancer. We studied the in vivo correlation of maspin expression with p53 mutation in malignant melanoma (MM) with and without use of tissue microarray (TMA). Seventy-seven MMs were immunostained on individual slides for maspin and p53 expression. Results were validated in 1 slide for each marker on a TMA system (TARP-2) with 498 tissue cores (0.6-mm diameter) from MM, other tumors, and normal tissue. The relationship between maspin and p53 in MM and carcinomas of other sites (breast, ovary, colon, lung, and prostate) was delineated using Pearson chi analysis. The inverse relationship between maspin and p53 expression predicted by hypothesized p53 regulation of maspin transcription, or any other correlation between these 2 markers, is not demonstrated in MM cases, using either classic individual slide (P=0.20) or TMA (P=0.85) methods when cutoffs for both markers are set at 10% or greater of cells staining. Even when cutoffs are altered with respect to either intensity or percentage of cells staining, no relationship is demonstrated between these markers, with either TMA or the conventional slide method. TMA immunostaining also showed no such relationship in carcinomas of the various other sites sampled-including breast and prostate, where previous studies have suggested a linkage. Despite published experimental evidence linking these 2 markers, this study failed to demonstrate correlation between maspin loss and p53 expression in MM using both individual slides and TMA, or in TMA of other carcinomas. Use of TMA is a quick, easy, and inexpensive method of immunohistochemical analysis of large numbers of cases, both to validate results obtained from individual slides and to assess specificity in a variety of neoplasms. However, heterogeneity and minimal tumor may lead to variable results.