WB
Western blot analysis of lysates from RAW 264.7 cells using IL1β Rabbit mAb at 1:1000 dilution incubated overnight at 4℃. Raw264.7 cells were treated by LPS (1μg/ml) at 37℃ for 8 hours. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 30 μg per lane. Blocking buffer: 3 % nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 90s.
ICC/IF
Confocal imaging of RAW 264.7 cells (treated with BFA and LPS) and RAW 264.7 cells (untreated) using IL1β Rabbit mAb (dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.