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All samples sintered at constant temperature at 1100oC for 2 hr.
The conjugation reactions were performed at 37°C for 2 hr.
Antibodies were bound for 2 hr at room temperature.
For cryosections, eyes were retrieved and immediately immersed in 4% paraformaldehyde for 2 hr.
The plates were then blocked in 1% BSA in TBST for 2 hr at 37°C.
Secondary antibodies were applied to the sections for 2 hr at room temperature.
Rabbit HRP-anti-Ddig antibody (Dako, 1∶100) was used for 2 hr.
After washing, the cells were cultured in RPMI with 10% FBS for 2 hr.
Primary antibody incubations were for 2 hr at room temperature or overnight at 4°C.
Membrane vesicles were collected by ultracentrifugation at 111,000×g for 2 hr at 4°C.
The permeabilized cells were incubated with primary antibodies in PBS for 2 hr at RT.
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