Immunofluorescent analysis of iNOS in A549 Cells (ab7910). Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a iNOS polyclonal antibody (ab3523) at a dilution of 1:20 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. iNOS staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

IHC with a indirect immunoalkaline phosphatase (APAAP) technique using ab3523. Shows iNOS expression within vessels walls in the Locus Coeruleus after death from septic shock. Picture kindly given to us by Prof. Djillali Annane.

All lanes : Anti-iNOS antibody (ab3523) at 1/3000 dilutionLane 1 : Whole tissue lysate prepared from mouse lungsLane 2 : Whole tissue lysate prepared from mouse lungsLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/10000 dilutiondeveloped using the ECL techniqueObserved band size : 131 kDa (why is the actual band size different from the predicted?)Exposure time : 30 secondsThis image is courtesy of an anonymous AbreviewSee Abreview

ab3523 staining iNOS in murine lung tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Tissue was fixed in paraformaldehyde, blocked with 5% BSA for 30 minutes and then incubated with ab3523 at a 1/50 dilution for 18 hours at 4°C. The secondary used was an undiluted HRP conjugated goat polyclonal.See Abreview

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human eart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing iNOS (ab3523) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human lung tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing iNOS (ab3523) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.