Alexa fluor 488 goat anti rabbit igg antibody conjugate

Induced E. coli cultures as described above were washed with PBS, and then 15 µl of cell suspension was air-dried on a glass coverslip before heat fixing. The coverslip was washed in PBS to remove unfixed cells before blocking in PBS containing 5% v/v foetal calf serum (FCS) for 1 h. After washing three times with PBS, coverslips were incubated with serotype appropriate rabbit anti-capsule antibody (Statens Serum Institut, Denmark) at a dilution of 1 : 1000 in PBS 5% FCS for 1 h. Following another three washes with PBS, the coverslips were incubated with Alexa Fluor 488 goat anti-rabbit IgG antibody conjugate (Life Technologies Ltd., UK) 1 : 1000 in PBS 5% FCS for 45 min. The coverslips were washed three times with PBS and allowed to air dry before mounting with SlowFade Gold antifade Mountant with DNA-specific counter stain 4′,6-diamidino-2-phenylindole (Life Technologies Ltd., UK). Each slide was visualized under fluorescence using an Olympus FluoView laser scanning microscope (Olympus Imaging and Audio Ltd.)