2-DIMENSIONAL IMMUNOBLOT ANALYSIS OF ALLERGENS OF CLADOSPORIUM-HERBARUM

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 analysi s revealed 10 human IgE-binding components with molecular masses rangi ng from 15 to 143 kDa. Within the same molecular mass range, five comp onents were detected under non-reduced conditions. Examination of the specificities of 10 individual sera revealed that two patients were se nsitive to almost all the allergens, whereas the majority reacted with only some components. Two-dimensional immunoblot was employed to furt her analyse the allergens in the extract. A silver-stain of the two-di mensional gel revealed that there were more than 30 different protein components visible in the extract. Immunoblot of the proteins separate d under denatured conditions disclosed that 17 spots were reactive wit h human atopic sera.