Identification of L. angustifolius seed storage proteins. Lupin flour proteins were separated by 2D-PAGE and (A) stained with Coomassie-blue stained or (B) blotted onto a membrane, which was probed with serum from lupin-allergic individuals, to identify potentially allergenic IgE-binding proteins. Protein spots that were either IgE-binding (spots 3 - 59, 94) or non-IgE-binding (spots 87 - 89, 97-114) were analysed by mass spectrometry, and those for which identifications were made are enclosed by ovals, with different colours corresponding to different proteins as shown in the figure. Sections of the gel and blot (boxes i, ii and iii) have been enlarged to show more detail, with Coomassie-blue stained gels on the top and IgE-binding proteins on the bottom panel for each section. In the enlarged boxes i, ii and iii spots that bind IgE are shown in black and those that do not in red. IgE binding was determined by aligning the original film and the Coomassie-blue stained gel but for 4 spots (37, 38, 51, 57) the resolution of the gel does not give a clear image of this binding. 'Beta?' indicates spots for which the form of conglutin β could not be determined.