Western Blot Analysis of Drosophila Seminal Proteins

Proteins were extracted from 3–4 day old virgin male AG or heads (as in [38 (link)]). Western blotting for Acp36DE, Ovulin, Acp62F, sex peptide (SP), and Actin was performed as previously described [39 (link)]. Western blotting for Rh1 was as previously described [40 (link)]. The Rh1 monoclonal antibody (4C5) was purchased from the Developmental Studies Hybridoma Bank (University of Iowa). The size markers in Fig. 2A and Fig. 3D are from the SeeBlue Plus2 Protein Standard (Invitrogen). All Western blot analyses were performed in triplicate independent experiment. Representative images are shown in the figures.