Two important Cladosporium herbarum allergens (Ag-54 and Ag-32) were partly purified by means of diafiltration, gel filtration and isoelectric focusing. Modifications of quantitative immunological techniques, including autoradiography, were used as assay systems for allergenic activity and to provide a basis for evaluation of the immunological purity of the allergens. Ag-54 appeared to have a molecular weight of approximately 25,000 daltons and an isoelectric point of 5.0. The molecular weight of Ag-32 appeared to be approximately 13,000 daltons. It consisted of 5 molecular species differing in isoelectric points from 3.4 to 4.4 in intervals of approximately 0.25 pH units. These variants of Ag-32 seemed to be antigenically and allergenically identical.