Fluorescein isothiocyanate fitc conjugate anti goat antibody

All the materials used were of the highest grade of purity and did not need further purification. Cyclophosphamide, acrolein, human tubular fluid (HTF) media, anti-α tubulin antibody, fluorescein isothiocyanate (FITC) conjugate anti-goat antibody, 4′,6-diamidino-2-phenylindole (DAPI), 1% BSA (Bovine Serum Albumin), 0.1% M Glycine, and 0.1% Triton X- 100 were obtained from Sigma-Aldrich (St. Louis, MO, USA). Normal Goat Serum (2%) was from Invitrogen (Grand Island, NY) and 0.2% Powder Milk from grocery. Metaphase II oocytes (with and without CC) from a B6C3F1 mouse crossed with a B6D2F1 mouse were obtained commercially (Embryotech Inc.) in cryopreserved straws using ethylene glycol-based slow freeze cryopreservation protocol. The oocyte spindle repolymerizes to normal structure when incubated in media for 60–120 min at 37 °C and 5% CO₂ prior to induction of oxidative stress [33 (link)]. This mechanism actually has helped support utilization of frozen oocytes for studying spindle damage. The use of frozen-thawed oocytes is well accepted as many studies have been published in the past utilizing frozen thawed oocytes and effects of oxidative stress on spindle morphology.