Figure 3

Silver staining of 2-DE separation (A) and typical 2-DE IgE-binding spectra of Cup.s . pollen proteins (B-D). First dimension (IEF) was carried out in a 2-11 pH gradient and the second dimension in an 8-18% acrylamide gradient gel. Molecular masses are expressed in kilodaltons (kDa); pI corresponds to isoelectric points. Representative IgE binding patterns B, C, and D correspond, respectively, to 2-DE immunoblotting allergen profiles of the patients 2, 18, and 20 of Figure 1 and Table 1. The pattern 3B is constituted of numerous IgE-reactive proteins with molecular masses ranging from 35 to 94 kDa and pI values distributed from 3.5 to 8.5. The patterns 3C and 3D revealed a very poor heterogeneity (IgE binding protein spots are shown by black arrows) and particularly recognized very basic spots of ~14 kDa.