Identification of the immunologic specificity of antinuclear antibodies (ANAs) in the various systemic rheumatic diseases has become increasingly important. The standard immunofluorescence technique may enable detection of antibodies to nuclear antigens present in abundance in the nucleus, such as DNA, histones, Sm, nuclear ribonucleoprotein (nRNP), and SS-B/La. The nuclear antigens present in low concentrations, such as SS-A, proliferating cell nuclear antigen (PCNA), rheumatoid arthritis nuclear antigen (RANA), and Ku antigens, are unique to cell types, and their detection requires special substrates or reagent systems. Anti-Sm, anti-Scl-70, anticentromere, and anti-PM-1 are characteristic serologic markers for systemic lupus erythematosus, scleroderma, the CREST syndrome of scleroderma, and polymyositis, respectively. Distinct profiles of ANA characterize different rheumatic diseases. A number of ANAs are found in SLE, whereas other diseases are characterized by the presence or absence of a certain ANA or by differences in mean ANA titers. Specific ANAs have been used to isolate and characterize nuclear antigens at molecular and functional levels.