Anti-iNOS antibody (ab15323)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - iNOS antibody (ab15323)

Human lung stained with ab15323 in IHC paraffin fixed sections.

  Immunohistochemistry (Frozen sections) - iNOS antibody (ab15323)

We performed immunostaining for the three NOS isoforms to examine their localization in mesenteric arteries of WT and the three genotypes of NOS-/- mice. In WT mice, the immunoreactivity of eNOS and nNOS, but not that of iNOS, was noted mainly in the endothelium, and in eNOS-/- and n/eNOS-/- mice, the expression of nNOS and iNOS was noted, respectively. In contrast, in n/i/eNOS-/- mice, none of the NOS isoforms were noted, as expected. Furthermore, we performed Western blot analysis to quantify the protein expression of NOS isoforms in the whole mesentery. In WT mice, the expression of both eNOS and nNOS was noted, whereas that of iNOS was minimal. In eNOS-/- mice, the expression of nNOS was again noted, and in n/eNOS-/- mice, the expression of iNOS was significantly up-regulated compared with WT or eNOS-/- mice. In n/i/eNOS-/- mice, none of the NOS isoforms were noted, as expected.

Image from Takaki A et al, J Exp Med 205:2053-63 (2008), Fig 4.

  Immunocytochemistry/ Immunofluorescence - iNOS antibody (ab15323)

ICC/IF image of ab15323 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab15323, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323)

ab15323 staining macrophages in mouse spleen sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/50 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Carl Hobbs

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323)

ab15323 staining rat liver (lower image) and spleen (upper image) sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/100 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Carl Hobbs

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