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. 2014 Jan 9;9(1):e82341. doi: 10.1371/journal.pone.0082341

Figure 1. Immunochemical cross-reactivity.

Figure 1

A) Detection of specific human serum IgE antibodies by indirect ELISA. Microtiter plates were coated with CMP, SP, recombinant α-subunit of β-conglycinin “α” or α-T, and sequentially incubated with sera of CMA patients and anti-human IgE specific conjugate. 15/15 sera contained IgE antibodies specific for CMP, 13/15 sera contained IgE antibodies specific for α and 14/15 sera contained IgE antibodies for α-T. B) SDS-PAGE and immunoblotting of CMP and SP. SDS-PAGE was performed under non reducing conditions for CMP (upper panel), SP (middle panel), and purified recombinant α and the α-T fraction (lower panel). Immunoblottings were developed with patient sera containing milk-specific IgE antibodies or control sera (from non-allergic patients) (1∶4), CMP-specific rabbit polyclonal antiserum (1∶1,000), and α-casein-specific monoclonal antibody (1D5 mAb 1∶1,500). CMP: Cow's milk protein, cas: caseins, SP: soybean protein, G: 11S glycinin subunits; β-conglycinin subunits are indicated as α, α′ and β. M: Molecular masses are given on the left in kilo Daltons (kDa).