iNOS Polyclonal Antibody for Western Blot, IF, ICC, IHC (F) and IHC (P)
iNOS Antibody (PA3-030A) in IF
Immunofluorescence analysis of iNOS was performed using 70% confluent log phase A549 cells treated with a combination of TNF-α (5 ng/ml), IFNγ (50 U/ml) and IL-1β (50 U/ml) for 18 h. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with iNOS Rabbit Polyclonal Antibody (PA3030A) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the untreated cells with no signal. Panel f represents the no primary control. The images were captured at 60X magnification.