Aedes aegypti eggshell purification. A) Eggshells purified by differential sedimentation from ovaries dissected at 72 hPBM and visualized by light microscopy. Top image, ovary dissected at 72 hPBM. Bottom images, eggshells purified by differential sedimentation. B) Ovary (Ov) and eggshell (ES) protein preparations dissolved in SDS/β-mercaptoethanol buffer resolved using 4–15% SDS-PAGE and stained with Coomassie blue R. The electrophoretic migrations of molecular weight markers (M - Precision Plus Protein Kaleidoscope, Bio-Rad Laboratories) and their corresponding molecular weights are indicated at the left of the figure. The excised gel slices submitted to mass spectrometry are indicated by letters A-J, and the identified proteins are listed in Additional file 1. Vitellin subunits are indicated with asterisks.