Mattress dust samples from 42 homes in Baltimore, Md., were analyzed by ELISA and rocket immunoelectrophoresis (RIE) for major excrement allergens (P1 [equal to Dp 42], Df 6, and Dm 6) of house dust mites, Dermatophagoides pteronyssinus, D. farinae, and D. microceras, by RIE for cat major epithelial allergen (cat Ag 1) and horse major epithelial antigen (horse Ag 11), and by counter-current immunoelectrophoresis (CCIE) for cat and dog antigens. Dp 42 was detected by ELISA in 24 samples (57%) and Df 6 in 39 samples (92%). All samples were negative to Dm 6. Recorded levels varied from 50 ng of allergen per gram of dust (detection limit) to 15,000 ng/gm. A combined content of Dp 42 and Df 6 greater than 700 ng/gm was associated with a positive reaction in RIE. Concentrations of Dp 42 and Df 6 did not correlate (rS = 0.21; p greater than 0.05). Cat and dog antigens were frequently detected by CCIE (77% and 63%, respectively). CCIE titer to cat correlated well with quantitation of cat Ag 1 by RIE (p less than 0.01). Horse antigen was detected in few homes (5%). None of the specific allergen determinations correlated with levels of human serum albumin determined by RIE.