Figure 3: Detection of immuno-reactive cricket proteins to patient sera: A representative western blot of crude protein extracts from different body parts of the insect (20 μg each) blotted with patient sera (lanes 6-9) showed more intense immuno-reactive bands as compared to lanes blotted with control sera (1-4) (A); lanes 1and 6, abdomen/thorax; lanes 2 and 7, guts; lanes 3 and 8, wings/legs; lanes 4 and 9, hearts (n=3). Protein extracted the most immunoreactive component of the insect (wings/legs, see A) was semipurified by a single step chromatography using Superdex 200 (B) or two-step chromatography using Superdex 200 followed by DEAE Sepharose (C) 1 μg fractions blotted with patient sera. The four prominent bands shown in Figure A are enriched upon purification despite the use of 20 fold less protein. Figure B and C are representative of 2 independent experiments.