Fig. 5.

Protein and allergen profile. (A) 40 µg raw shrimp extract, 8 µg boiled shrimp extract, and 2 µg recombinant tropomyosin (rTM) were resolved on 10% SDS-PAGE. Note the significant loss of proteins in the boiled preparation. Immunoblot was performed with serum pool (n = 8) from PBS controls, rTM-, raw extract- and boiled-extract sensitized and challenge mice against (B) rTM (the 46 kDa protein band of his-tagged tropomyosin expressed in pET30a), (C) raw shrimp extract and (D) boiled shrimp extract. Note that IgE binding was only detected against tropomyosin in all experimental groups. (E) Pool of sera from raw extract-sensitized mice with higher IgE level to raw extract than to rTM (n = 5) were incubated against raw extract, and identified other shrimp allergens including hemocyanin (80 kDa) and glycogen phosphorylase (100 kDa). L, protein marker (New England Biolabs) with reference molecular weight (kDa) indicated