Goat anti rabbit igg as1109

Frozen tissue sections and cells grown on glass-coverslips were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 for 10 min, and blocked in 10% normal goat serum for 1 h. The anti-CXCL12/SDF1 rabbit polyclonal antibody (Cat#: AF6612, Beyotime Biotechnology, 1:100) was diluted in blocking buffer and placed on the cells at 4°C overnight. After the incubation, slides were washed three times in PBS and then incubated with goat anti-rabbit IgG (AS1109; Aspen Biotechnology, Wuhan, China; 1:100) for 1 h at 37 °C. For filamentous actin (F-actin) staining, cells on coverslips were incubated with FITC-phalloidin (Cat#: C1033, actin-tracker green, Beyotime Biotechnology) in PBS for 2 h at room temperature. Nuclei were stained using 4′6′-diamidino-2-phenylindole (DAPI) solution for 10 min at room temperature. Imaging was performed using an FV1200 (Olympus Corporation) confocal microscope.

Primary melanocytes (5×10⁵) were seeded in 6-well culture plates containing coverslips. After attachment, the cells were treated with multiple UVB exposures for a cumulative dose of 70 mJ/cm² (8.75 mJ/cm² per exposure in 5 h intervals for a total of 8 times). Subsequently, the cells were immediately fixed in 4% paraformaldehyde in PBS for 30 min at room temperature, permeabilized with 0.3% Triton X- 100 in PBS for 15 min and then blocked for 1 h at 37°C using a blocking buffer containing 10% normal goat serum (cat. no. C0265; Beyotime Institute of Biotechnology). The anti-MMP9 (cat. no. ab76003; Abcam; 1:200) antibody was diluted in blocking buffer and placed on the cells at 4°C overnight. After the incubation, cells on coverslips were washed three times in PBS and then incubated with goat anti-rabbit IgG (AS1109; Aspen Biological; 1:50) for 1 h at 37°C. Nuclei were stained using 4′6′-diamidino-2-phenylin-dole (DAPI) solution for 10 min at room temperature. Imaging was performed using an FV1200 (Olympus Corporation) confocal microscope.