Porcine albumin

Since we anticipated identifying the greatest number of proteins from ovaries during the trophic phase of oogenesis (24 hr PBM), we chose to quantitate differences in ovary protein expression based on iron content of the diet at this developmental stage by labeling sample peptides using a tandem mass tag (TMT) Isobaric and Isotopic Mass Tagging kit. Mosquitoes were maintained as described above. Mated, female mosquitoes were fed one of two isoproteinic diets or a Porcine bloodmeal (Porcine BM, 603 ng Fe/μl) maintained at 27°C in glass feeders for 2 hr. The isoproteinic diets were: Kogan’s (1990) (link) artificial bloodmeal (ABM) with hemoglobin ((+)Fe): 0.8% (w/v) Porcine Hemoglobin (Sigma, St. Louis, MO); 1.5% (w/v) Porcine IgG (Sigma); 10% (w/v) Porcine Albumin (Sigma); and 5 mM ATP (Sigma) in feeding buffer (~56 ng Fe/μl) and Kogan’s (1990) (link) ABM without hemoglobin ((-)Fe): 10.7% (w/v) Porcine Albumin; 1.6% (w/v) Porcine IgG; and 5 mM ATP in feeding buffer (~24 ng Fe/μl). Tissues were collected as described above for early stage of oogenesis (24 hr PBM). Ovary proteins were extracted, and protein concentrations were determined as described above. Samples were flash frozen in liquid nitrogen and stored at −80°C until TMT-labeling.