WB
Western blot analysis of nNOS (Phospho-S1417) expression in A549 (A), HEK293T (B) whole cell lysates.
IHC
Immunohistochemical analysis of nNOS (Phospho-S1417) staining in human stomach cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
ICC/IF
Immunofluorescent analysis of nNOS (Phospho-S1417)
staining in HepG2 cells. Formalin-fixed cells were
permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes
and blocked with 3% BSA-PBS for 30 minutes at room
temperature. Cells were probed with the primary antibody in
3% BSA-PBS and incubated overnight at 4 °C in a hidified
chamber. Cells were washed with PBST and incubated with a
AF488-conjugated secondary antibody (green) in PBS at room
temperature in the dark. Phalloidin - AF594 was used to stain
Actin filaments (red). DAPI was used to stain the cell nuclei
(blue).