“…A human mAb against Api m 1 (BVA2) was used to characterize further the Api m [1/2] fusion protein. 16 Immunoblotting For Western blot analysis, equimolar amounts (1.5 mmol/L) of the 3 recombinant proteins, Api m 1, Api m 2, and Api m [1/2], were electrophoresed by 4% to 12% Bis-Tris polyacrylamide NuPAGE gels (Invitrogen, Basel, Switzerland) under denaturing, reducing conditions and transferred to nitrocellulose membrane (Amersham, Little Chalfont, United Kingdom). 17 The binding of IgE in pooled serum from 4 individuals allergic to BV was detected with rabbit antihuman IgE (Dako Inc, Burlingame, Calif) and developed by goat antirabbit biotin conjugated IgG antibody (Dako Inc) and streptavidin-horseradish peroxidase (Sigma Chemical Co, St Louis, Mo).…”