Alexa fluor 488 monoclonal antibody labeling kit

The anti-RSV monoclonal antibody was labeled with Alexa Fluor 488, and the anti-Vg monoclonal antibody was labeled with Alexa Fluor 594 according to the Alexa Fluor 488 Monoclonal Antibody Labeling Kit and Alexa Fluor 594 Protein Labeling Kit (Invitrogen) instructions, respectively. The ovaries were dissected from L. striatellus nymphs and female adults at four stages: 12 h postemergence (Adult I), 24 h postemergence (Adult II), 36 h postemergence (Adult III), and 48 h postemergence (Adult IV) in cold distilled water in a glass plate, and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 2 h at room temperature. The fixed ovaries were incubated in osmotic buffer (2% triton v/v in PBS) for 4 h at room temperature, and then incubated with labeled anti-RSV and anti-Vg antibody for 2 h at room temperature. The samples were viewed with a laser scanning confocal microscope (Leica TCS SP5) and the images saved by Leica LAS AF Lite.