2.12. Immunofluorescence
SECONDARY, FLUOR
Slides were washed with phosphate-buffered saline (PBS) and fixed with 4% paraformaldehyde for 30 min and measured with 0.2% Triton-X 100 for 10 min. Then, they were blocked with 5% bovine serum albumin and incubated with Alexa Fluor 555-antimouse IgG (H + L) (Cell Signaling Technology, MA, USA) for 1 hr at room temperature. For the staining of complement 3 (C3), frozen sections were incubated with rabbit antimouse C3 (Abcam, MA, USA) followed by the staining of secondary Alexa Fluor 488-antirabbit IgG (H + L) (Proteintech, Wuhan, China). After three PBS washes, the slides were counterstained with DAPI for 1 min and then observed under the Confocal Laser Scanning Microscope FV3000 (Olympus, Tokyo, Japan).