Camp antibody

Cells were seeded on the glass coverslips in 24-well plates. After washing with PBS three times, cells were fixed with 2% paraformaldehyde for 15 min at room temperature followed by permeabilization with 0.1% Triton × −100. Afterwards, 3% bovine serum albumin (BSA) was used to block the unspecific binding sites for 50 min at room temperature. Cells were then incubated overnight at 4 °C with the primary antibody: 1:100 diluted anti-CD68 antibody (Abcam, MA, USA), 1:50 diluted iNOS antibody (Santa Cruz Biotechnology, CA, USA), 1:50 diluted ARG-1 antibody (Santa Cruz Biotechnology), 1:50 diluted CAMP antibody (Abcam). Samples were then incubated with secondary antibody: 1:200 diluted Donkey Anti-Rabbit IgG H&L, Alexa Fluor^® 594 (Abcam), 1:200 diluted Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594 (Abcam) for 50 min at room temperature. After being rinsed with PBS, 4’, 6-diamidino-2-phenylindole (DAPI) was added to stain the cell nucleus and samples were visualized under immunofluorescence microscopy (Nikon, Eclipse 80i, Tokyo, Japan).