Identification of dimeric Bet v 1 during WT protein preparation.
a, separation of monomeric and dimeric Bet v 1 via ion exchange (top panel, peak corresponding to dimer indicated by black lines) and size exclusion chromatography (bottom panel). b, analysis of size exclusion chromatography fractions via native (top panel) and SDS (middle and bottom panels) PAGE. Comparison of reducing (middle panel) and nonreducing (bottom panel) SDS-PAGE demonstrates the reduction sensitivity of the dimer. c, SDS- and clear native-PAGE revealed stability of the correctly folded dimer under reducing conditions (bottom panel, right side). The bands corresponding to dimeric Bet v 1 are boxed. Lane M, marker; lanes 17 and 35, molecular mass of the marker band in kDa.