Two-dimensional immunoblot analysis of allergens of Cladosporium herbarum

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Abstract
In the present study, allergens of Cladosporium herbarum were analysed by SDS-PAGE, two-dimensional gel electrophoresis and immunoblot. The SDS-PAGE under reduced conditions followed by western blotting analysis revealed 10 human IgE-binding components with molecular masses ranging from 15 to 143 kDa. Within the same molecular mass range, five components were detected under non-reduced conditions. Examination of the specificities of 10 individual sera revealed that two patients were sensitive to almost all the allergens, whereas the majority reacted with only some components. Two-dimensional immunoblot was employed to further analyse the allergens in the extract. A silver-stain of the two-dimensional gel revealed that there were more than 30 different protein components visible in the extract. Immunoblot of the proteins separated under denatured conditions disclosed that 17 spots were reactive with human atopic sera.