Figure 3

Blocking western blot. Maternal plasma samples incubated without any target antigen (U–unblocked) or blocked with lactate dehydrogenase (LDH), collapsin response mediator protein (CRMP) 1, stress-induced phosphoprotein 1 (STIP1; mother with reactivity to all three proteins), CRMP2 (mother who is CRMP1− and CRMP2 +), Y-box-binding protein (YBX1) and Cypin (mother with reactivity to Cypin, STIP1 and CRMP1) were used to probe a western blot containing Rhesus macaque fetal brain. The pure recombinant proteins were incubated with the plasma sample diluted 1:400 overnight, centrifuged and applied to the membrane blot containing fetal monkey brain. Arrowheads indicate location of the target antigen removed by overnight pre-incubation. Note that only the specific band(s) is/are removed following pre-incubation with each individual target protein indicating no cross-reactivity between the candidate antigens.