Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cells treated with LPS (0.1mg/ml), for 6 hours labeling iNOS with ab178945 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).Confocal image showing cytoplasmic staining on RAW 264.7 cells, treated with LPS (0.1mg/ml), for 6 hours.The nuclear counterstain is DAPI (blue).Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:--ve control 1: ab178945 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
![All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/20000 dilutionLane 1 : Untreated RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysateLane 2 : RAW 264.7 whole cell lysate treated with 0.1 mg/ml LPS for 6 hoursLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/8682_ab178945-245976-ab178945WBa.jpg)
All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/20000 dilutionLane 1 : Untreated RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysateLane 2 : RAW 264.7 whole cell lysate treated with 0.1 mg/ml LPS for 6 hoursLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
![All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/1000 dilutionLane 1 : Human fetal liver lysateLane 2 : Human fetal brain lysateLane 3 : Human fetal spleen lysateLane 4 : Human fetal lung lysateLysates/proteins at 20 µg per lane.SecondaryAnti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/8683_ab178945-245975-ab178945WBb.jpg)
All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/1000 dilutionLane 1 : Human fetal liver lysateLane 2 : Human fetal brain lysateLane 3 : Human fetal spleen lysateLane 4 : Human fetal lung lysateLysates/proteins at 20 µg per lane.SecondaryAnti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
iNOS was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate treated with 1µg/ml LPS for 24h with ab178945 at 1/100 dilution.Lane 1: RAW 264.7 whole cell lysate treated with 1µg/ml LPS for 24h,10ug (Input).Lane 2: ab178945 IP in RAW 264.7 hole cell lysate treated with 1µg/ml LPS for 24h.Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab178945 in RAW 264.7 whole cell lysate reated with 1µg/ml LPS for 24h.Western blot was performed from the immunoprecipitate using ab178945 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/1500.Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 30 seconds.